Applications of protein VdAL in improving output, product quality and drought resistance of plant and in improving fruit coloring of plant
10806145 ยท 2020-10-20
Assignee
Inventors
- Junsheng Qi (Beijing, CN)
- Zhizhong Gong (Beijing, CN)
- Xuhui Hong (Beijing, CN)
- Dingpeng Zhang (Beijing, CN)
Cpc classification
A01N25/00
HUMAN NECESSITIES
C12N15/8245
CHEMISTRY; METALLURGY
Y02A40/146
GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
C12N15/8242
CHEMISTRY; METALLURGY
C12N15/8261
CHEMISTRY; METALLURGY
C12N15/8249
CHEMISTRY; METALLURGY
C12N15/8243
CHEMISTRY; METALLURGY
A01N25/00
HUMAN NECESSITIES
International classification
C12N15/82
CHEMISTRY; METALLURGY
A01N63/10
HUMAN NECESSITIES
Abstract
Provided are applications of protein VdAL in improving the output, product quality and drought resistance of a plant and in improving fruit coloring of the plant. The protein VdAL is selected from one of A1) to A3): A1), protein having an amino acid sequence that is 1; A2) protein that is derived from A), that has same functions and that is obtained by substituting and/or deleting and/or adding one or several amino acid residues for, from or to the sequence 1; and A3) fusion protein obtained by connecting labels to an N end and/or a C end of A1) or A2).
Claims
1. A method of treating a plant with a biological agent, the method comprises applying a biological agent to a plant, wherein said biological agent comprises a protein comprising the amino acid sequence as set forth in SEQ ID NO: 1, wherein said protein has VdAL protein activity in said biological agent, and wherein said application of said biological agent to the plant increases plant yield, increases plant growth, improves plant product quality, improves drought resistance or promotes fruit coloration of said treated plant as compared to a control plant of the same species that is not treated with said biological agent and is grown under identical conditions.
2. The method of claim 1, wherein said protein is a fusion protein comprising a labeled peptide tag at N-terminal or C-terminal end of said protein as set forth in SEQ ID NO: 1, and wherein said fusion protein has VdAL protein activity of said SEQ ID NO: 1.
3. The method according to claim 1, wherein said plant is a plant of dicotyledoneae or monocotyledonae.
4. The method according to claim 1, wherein said plant growth is vegetative growth and/or reproductive growth.
5. The method according to claim 1, wherein said plant product quality is nutrient content or trace elements.
Description
BRIEF DESCRIPTION OF THE DRAWINGS
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(19) In the context, 1/300, 1/600, 1/900, 1/1000, 1/200, 1/250, 1/3000, 1/4000, 1/5000, 1/6000, 1/7500, 1/10000, 1/20000 and 1/40000 times VdAL respectively represent 300 times diluted VdAL, 600 times diluted VdAL, 900 times diluted VdAL, 1000 times diluted VdAL, 2000 times diluted VdAL, 2500 times diluted VdAL, 3000 times diluted VdAL, 4000 times diluted VdAL, 5000 times diluted VdAL, 6000 times diluted VdAL, 7500 times diluted VdAL, 10000 times diluted VdAL, 20000 times diluted VdAL, and 40000 times diluted VdAL.
DETAILED DESCRIPTION OF THE EMBODIMENTS
(20) The present invention will be described in further detail with reference to specific examples, which are provided for illustration of the present invention only, but not intended to limit the scope of the present invention.
(21) Experimental methods indicated in the following examples are conventional approaches, unless otherwise specified.
(22) Materials, agents and the like used in the following examples are all commercially available, unless otherwise specified.
(23) Escherichia coli JM109 in the following examples is a product of Beijing Solarbio Science & Technology Co., Ltd. The catalog number thereof is C1300.
(24) Vector pET42a (+) in the following examples is a product of Beijing SinaSun Biotechnology Co., Ltd. The catalog number thereof is S 18-16.
(25) Xinluzao No. 58 in the following examples is a product of Xinjiang Jinmian Seed Industry Science & Technology Co., Ltd. In addition, Lu No. 30, 616, Lu No. 25 and Luyanmian No. 24 are products of Shandong Nongxing Seed Industry Co., Ltd.
(26) Variety of watermelon, Jinlidu, in the following examples is a product of Shouguang Jishan Commercial and Trading Co., Ltd.
(27) Zhongmai 816 in the following examples is a product of Beijing Longshengyuan Science & Technology Development Co., Ltd.
(28) Variety of tomato, Fux, in the following examples is a product of Shouguang Jinpeng Seed Industry Co., Ltd.
(29) Variety of green-stem cabbage, Huali FI, in the following examples is a product of Shanghai Hongqiao Tianlong Seed Industry Co., Ltd.
(30) Variety of cucumber, Jinza No. 1, in the following examples is a product of Shandong Nongxing Seed Industry Co., Ltd.
(31) Variety of cherry tomato, Jingdan No. 2, in the following examples is a product of Beijing Beinong Lvheng Science & Technology Development Co., Ltd.
(32) Variety of strawberry, Fengxiang, in the following examples is a product of Beijing Beinong Luheng Sci-Tech Development Co., Ltd.
(33) Variety of melon, Super Cuibaoxiang, in the following examples is a product of Jinan Ruimao Agricultural Science & Technology Development Co., Ltd.
(34) Variety of sweet pepper, Ruiyou 816, in the following examples is a product of Jinan Ruimao Agricultural Science & Technology Development Co., Ltd.
(35) Variety of soybean, King of China soybean, in the following examples is a product of Jinan Ruimao Agricultural Science & Technology Development Co., Ltd.
(36) Variety of rice, Nongda 502, in the following examples is a product of Beijing Beinong Luheng Sci-Tech Development Co., Ltd.
Example 1 Preparation of the Agent of VdAL
(37) The agent of VdAL was developed by China Agricultural University, and produced by Shandong Huimin Huhao Biotechnology Co., Ltd.
(38) 1. Construction of Recombinant Vector and Recombinant Bacterium
(39) The DNA molecule represented by nucleotides 1st to 894th of SEQ ID No. 2 in the Sequence Listing, i.e. the VdAL gene, was artificially synthesized. A sequence between NdeI and KpnI recognition sites of a vector pET42a (+) was replaced with the DNA molecule represented by nucleotides 1.sup.st to 894.sup.th of SEQ ID No. 2 in the Sequence Listing, i.e., the VdAL gene, while other sequences of the pET42a (+) remained unchanged. A recombinant vector was thus obtained. The obtained recombinant vector was named pET42a-VdAL. The recombinant vector pET42a-VdAL expresses the VdAL protein shown in SEQ ID No. 1 in the Sequence Listing.
(40) SEQ ID No. 2 is composed of 894 nucleotides encoding the amino acid sequence shown in SEQ ID No. 1.
(41) The pET42a-VdAL was transformed into Escherichia coli JM109 to obtain recombinant bacterium. The obtained recombinant bacterium was named JM109-pET42a-VdAL. JM109-pET42a-VdAL expresses the protein shown in SEQ ID No. 1.
(42) 2. Preparation of the Agent of VdAL
(43) JM109-pET42a-VdAL was fermented at 37 C. until OD value was 0.6, so that a pre-fermented liquid was obtained. IPTG was added into the pre-fermented liquid at the concentration of 1 mM to obtain induced liquid. The induced liquid was fermented for 6 hours at 25 C. to obtain fermented liquid. The fermented liquid was centrifuged. The supernate was removed. The obtained thalli pellet was disrupted. Then a dry powder agent (A in
(44) The VdAL in the agent of VdAL was detected by western-blot method. The primary antibody was anti-VdAL antibody which was polyclonal antibody obtained by using the protein shown in SEQ ID No. 2 as an immunogen. The result showed that the agent of VdAL comprised the VdAL protein (B in
(45) The agent of VdAL was dissolved into clear water to obtain liquids having the VdAL protein at the concentrations of 10 g/3 kg, 10 g/6 kg, 10 g/9 kg, 1 g/kg, 1 g/2 kg, 1 g/2.5 kg, 1 g/3 kg, 1 g/4 kg, 1 g/5 kg, 1 g/6 kg, 1 g/7.5 kg, 1 g/10 kg, 1 g/20 kg and 1 g/40 kg, respectively. These liquids were respectively named as 300 times diluted VdAL, 600 times diluted VdAL, 900 times diluted VdAL, 1000 times diluted VdAL, 2000 times diluted VdAL, 2500 times diluted VdAL, 3000 times diluted VdAL, 4000 times diluted VdAL, 5000 times diluted VdAL, 6000 times diluted VdAL, 7500 times diluted VdAL, 10000 times diluted VdAL, 20000 times diluted VdAL and 40000 times diluted VdAL.
Example 2 the Agent of VdAL can Promote the Growth of Cotton
(46) 1. The Agent of VdAL can Promote Germination of Cotton Seeds
(47) The seeds of variety, Xinluzao No. 58, were soaked into 5000 times diluted VdAL, 7500 times diluted VdAL and 10000 times diluted VdAL in Example 1 and clear water for 24 hours, respectively. Germination experiment was performed in big flower pots. The temperature was 25 C. The germination rate of cotton seeds was statistically analyzed in the 8.sup.th day after seeding. 100 seeds were treated in every treatment. The experiment was repeated three times.
(48) The results showed that the average germination rates of cotton were 74.00%, 75.50%, 71.00% and 70.23%, respectively, after the cotton seeds were treated with the 5000 times diluted VdAL, the 7500 times diluted VdAL, the 10000-times diluted VdAL and clear water. The average germination rates of cotton after the cotton seeds were treated with the 5000 times diluted VdAL, the 7500 times diluted VdAL and the 10000 times diluted VdAL are respectively 1.05 times, 1.08 times and 1.01 times that after the cotton seeds were treated with clear water. This indicated that the agent of VdAL could improve the germination rate of cotton seeds, and its capability of improving the germination rate of cotton seeds was changed with the change of the concentration of the agent of VdAL.
(49) 2. The Agent of VdAL can Improve the Yield of Cotton
Experiment I
(50) Field experiment was designed as follows. The experiment adopted randomized block design. Three replicate blocks were provided. Each replicate block was randomly provided with nine treatment areas which were respectively VdAL treatment area I of Lu No. 30, VdAL treatment area II of Lu No. 30, contrast treatment area of Lu No. 30, VdAL treatment area I of 616. VdAL treatment area II of 616, contrast treatment area of 616, VdAL treatment area I of Lu No. 25, VdAL treatment area II of Lu No. 25 and contrast treatment area of Lu No. 25. The area of each treatment area was 0.2 mu.
(51) At seedling stage, and blossoming and boll-forming early stage, Lu No. 30 in VdAL treatment area I of Lu No. 30 was respectively treated according to the following method. A 3000 times diluted VdAL of Example 1 with 45-50 kg per mu was sprayed to cotton leaves by the means of top spraying (mechanical spraying) to obtain Lu No. 30 treated with the 3000 times diluted VdAL. It was sprayed once at seedling stage, and blossoming and boll-forming early stage respectively. The spraying was performed on morning or at evening, and preferably not at the high temperature period at noon. The agent of VdAL was prohibited to be mixed with pesticides and fertilizers at spraying. The agent of VdAL should be sprayed once again if it rained within 2 hours after being sprayed.
(52) According to the treatment method of Lu No. 30 of VdAL treatment area I, the 3000 times diluted VdAL was replaced with a 5000 times diluted VdAL to treat Lu No. 30 of VdAL treatment area II. Other steps remained unchanged. Thus Lu No. 30 treated with the 5000 times diluted VdAL was obtained.
(53) According to the treatment method of Lu No. 30 of VdAL treatment area I, the 3000 times diluted VdAL was replaced with clear water to treat Lu No. 30 of contrast treatment area. Other steps remained unchanged. Untreated Lu No. 30 was obtained.
(54) According to the treatment method of Lu No. 30 of VdAL treatment area I, 616 of VdAL treatment area I and Lu No. 25 of VdAL treatment area I were respectively treated. Other steps remained unchanged. 616 and Lu No. 25 treated with the 3000 times diluted VdAL were obtained, respectively.
(55) According to the treatment method of Lu No. 30 of VdAL treatment area I, the 3000 times diluted VdAL was replaced with a 5000 times diluted VdAL to treat 616 of VdAL treatment area II and Lu No. 25 of VdAL treatment area II, respectively. Other steps remained unchanged. 616 and Lu No. 25 treated with the 5000 times diluted VdAL were obtained, respectively.
(56) According to the treatment method of Lu No. 30 of VdAL treatment area I, the 3000 times diluted VdAL was replaced with clear water to treat 616 of contrast treatment area and Lu No. 25 of contrast treatment area respectively. Other steps remained unchanged. Untreated 616 and Untreated Lu No. 25 were obtained, respectively.
(57) The cotton yield before first frost, cotton yield after first frost and total yield, of Lu No. 30 treated with the 3000 times diluted VdAL, Lu No. 30 treated with the 5000 times diluted VdAL, untreated Lu No. 30, 616 treated with the 3000 times diluted VdAL, 616 treated with the 5000 times diluted VdAL, untreated 616. Lu No. 25 treated with the 3000 times diluted VdAL, Lu No. 25 treated with the 5000 times diluted VdAL and untreated Lu No. 25 were statistically analyzed, respectively. The results were shown in Table 2.
(58) TABLE-US-00002 TABLE 2 The average yield of cotton in different treatments Cotton yield before Cotton yield first frost after first frost Total yield Yield Increasing Yield Total Increasing Variety Treatment (kg/mu) rate (%) (kg/mu) Yield rate (%) Lu Treatment 1 211.36 19.48 4.31 215.67 17.04 No. 30 Treatment 2 214.50 21.25 6.63 221.13 20.00 CK 176.90 7.37 184.28 616 Treatment 1 150.33 2.65 4.65 154.98 3.15 Treatment 2 183.86 19.06 7.66 191.52 19.69 CK 154.42 5.60 160.02 Lu Treatment 1 157.05 1.05 5.70 162.75 0 No. 25 Treatment 2 180.87 16.37 10.53 191.39 17.60 CK 155.43 7.32 162.75 Note: Treatment 1 represented the treatment with the 3000 times diluted VdAL, Treatment 2 represented the treatment with the 5000 times diluted VdAL, and CK represented untreatment (i.e., treatment with clear water).
(59) The results showed that the VdAL with a proper concentration could improve the yield of cotton. The cotton yield before first frost was improved by 19.48% and the total yield was improved by 17.04%, after Lu No. 30 was treated with the 3000 times diluted VdAL. The cotton yield of Lu No. 30 before first frost was improved by 21.25% and the total yield thereof was improved by 20.00%, the cotton yield of 616 before first frost was improved by 19.06% and the total yield thereof was improved by 19.69%, and the cotton yield of Lu No. 25 before first frost was improved by 16.37% and the total yield thereof was improved by 17.60%, after cotton was treated with the 5000 times diluted VdAL.
Experiment II
(60) Field experiment was designed as follows. The experiment adopted randomized block design. Three replicate blocks were provided. Each replicate block was randomly provided with four treatment areas which were respectively VdAL treatment area I, VdAL treatment area II, VdAL treatment area III and contrast treatment area. The area of each treatment area was 10 mu.
(61) At seedling stage, and blossoming and boll-forming early stage, Luyanmian No. 24 in VdAL treatment area I was respectively treated according to the following method. A 3000 times diluted VdAL of Example 1 with 45-50 kg per mu was sprayed to cotton leaves by the means of top spraying (mechanical spraying) to obtain Luyanmian No. 24 treated with the 3000 times diluted VdAL. The operation was performed at the speed of about 6 kilometers per hour. The agent of VdAL was sprayed once at seedling stage, and blossoming and boll-forming early stage respectively. The spraying was performed on morning or at evening, and preferably not at the high temperature period at noon. The agent of VdAL was prohibited to be mixed with pesticides and fertilizers at spraying. The agent of VdAL should be sprayed once again if it rained within 2 hours after being sprayed.
(62) According to the treatment method of VdAL treatment area I, the 3000 times diluted VdAL was replaced with a 5000 times diluted VdAL to treat VdAL treatment area II. Other steps remained unchanged. Luyanmian No. 24 treated with the 5000 times diluted VdAL was obtained.
(63) According to the treatment method of VdAL treatment area I, the 3000 times diluted VdAL was replaced with a 10000 times diluted VdAL to treat VdAL treatment area III. Other steps remained unchanged. Luyanmian No. 24 treated with the 10000 times diluted VdAL was obtained.
(64) According to the treatment method of VdAL treatment area I, the 3000 times diluted VdAL was replaced with clear water to treat contrast treatment area. Other steps remained unchanged. Untreated Luyanmian No. 24 was obtained.
(65) Plant number per mu, the number of boll setting of individual plant, total number of the boll per mu, the weight of single boll, ginning outturn and the yield per mu, of Luyanmian No. 24 in each treatment were statistically analyzed. Means of each item and increasing rates compared with the contrast were shown in Table 3 and Table 4.
(66) TABLE-US-00003 TABLE 3 Means and increasing rates of each item of Luyanmian No. 24 in each treatment Boll setting of individual plant Per mu plants the number of Total bolls per mu Plant boll setting of Total number Items number per Increasing individual Increasing of the boll per Increasing Treatment mu rate (%) plant rate (%) mu rate(%) Treatment 1 7901 2.2 9.2 +4.5 72283 +1.98 Treatment 2 8184 +1.4 9.1 +3.41 74006 +4.41 Treatment 3 8154 +1.0 8.7 1.14 71011 +0.18 CK 8072 / 8.8 / 70880 / Note: Treatment 1 represented the treatment with the 3000 times diluted VdAL, Treatment 2 represented the treatment with the 5000 times diluted VdAL, Treatment 3 represented the treatment with the 10000 times diluted VdAL, and CK representsuntreated (i.e., treated with clear water).
(67) TABLE-US-00004 TABLE 4 Means and increasing rates of each item of Luyanmian No. 24 in each treatment Weight of single boll Ginning outturn Weight of Increasing Ginning Yield per mu Items single boll rate outturn Increasing Yield per mu Increasing rate Treatment (g) (%) (%) rate (%) (kg/mu) (%) Treatment 1 5.4 0 41.2 0.24 392 +1.76 Treatment 2 5.5 +1.9 41.3 0.49 405.3 +5.22 Treatment 3 5.4 0 41.2 0.24 384.4 0.21 CK 5.4 / 41.1 / 385.2 / Note: Treatment 1 represented the treatment with the 3000 times diluted VdAL, Treatment 2 represented the treatment with the 5000 times diluted VdAL, Treatment 3 represented the treatment with the 10000 times diluted VdAL, and CK represented untreatment (i.e., the treatment with clear water).
(68) The results showed that the VdAL with a proper concentration could improve the number of plant per mu, the number of boll setting of individual plant, total number of boll per mu, the weight of single boll, ginning outturn and the yield per mu of cotton. The number of boll setting of individual plant, total number of boll per mu, ginning outturn and the yield per mu were increased, and the increasing rates were 4.5%, 1.98%, 0.24% and 1.76%, respectively, after cotton was treated with the 3000 times diluted VdAL. The number of plant per mu, the number of boll setting of individual plant, total number of boll per mu, the weight of single boll, ginning outturn and the yield per mu were increased, and the increasing rates were 1.4%, 3.41%, 4.41%, 1.9%, 0.49% and 5.22% respectively, after cotton was treated with the 5000 times diluted VdAL. The increasing rates of each item, when being treated with the 10000 times diluted VdAL, was lower than that of being treated with the 3000 times diluted VdAL and the 5000 times diluted VdAL. It indicated that the quality and the yield of cotton could be improved by applying VdAL with a proper concentration.
Example 3 the Agent of VdAL can Improve the Quality and Yield of Watermelon
(69) Field experiment was designed as follows. The experiment adopted randomized block design. Three replicate blocks were provided. Each replicate block was randomly provided with two treatment areas, which were respectively VdAL treatment area and contrast treatment area. The area of each treatment area was 0.2 mu.
(70) The variety of watermelon, Jinlidu, in VdAL treatment area was treated according to the following method. A 2000 times diluted VdAL of Example 1 with 30 kg per mu was sprayed to watermelon leaves by the means of top spraying (mechanical spraying) to obtain watermelon treated with the 2000 times diluted VdAL. It was sprayed at the seedling stage of watermelon for the first time, and sprayed once more 15 days after the first time. It was sprayed two times in total. The spraying was performed on morning or at evening, and preferably not at the high temperature period at noon. The agent of VdAL was preferably not mixed with pesticides and fertilizers at spraying. The agent of VdAL should be sprayed once again if it rained within 2 hours after being sprayed.
(71) According to the treatment method of VdAL treatment area, the 2000 times diluted VdAL was replaced with clear water to treat the contrast treatment area. Other steps remained unchanged. Untreated watermelons were obtained.
(72) The results showed that the agent of VdAL had a promoting effect on vegetative growth and fruit setting of watermelon (
(73) The water content, dry matter content, Vc content, soluble sugar content, titratable acid content, soluble solid content, peel thickness, protein content and coarse fiber content (Table 5), as well as the content of trace elements Ca, Cu, Fe, K, Mg, Mn, Na, P and Zn (Table 6), of watermelon with different treatments were respectively determined at the maturation stage of watermelon. The water content and the dry matter content (the percentage of dry matters in watermelon to fresh watermelon) were determined after the watermelon was dried. The Vc content was determined by a 2,6-dichlorophenol indophenol titration method. The soluble sugar content was determined by anthrone colorimetry. The titratable acid content was determined by colorimetry. The soluble solid content and the protein content were determined by spectrophotometry. The coarse fiber content was determined by the determination of crude fiber by Weende method. The content of trace elements Ca, Cu, Fe, K, Mg, Mn, Na, P and Zn were determined in each case by using trace element analyzer.
(74) TABLE-US-00005 TABLE 5 Average content of each index of watermelon in different treatments Index Untreated Treated by the VdLA Water content (%) 90.22 91.43 Dry matter content (%) 9.78 8.57 Vc(mg/100 g) 12.1 18.2 Soluble sugar (%) 6.83 7.44 Titratable acid (%) 0.09 0.09 Soluble solid (%) 9.1 9.1 Peel thickness (cm) 0.97 1.23 Protein (% DW) 5.54 6.33 Protein(% FW) 0.54 0.54 Coarse fiber(% DW) 2.06 2.02 Coarse fiber(% FW) 0.2 0.17
(75) TABLE-US-00006 TABLE 6 Average content of each trace element of watermelon in different treatments No. Index Untreated treated by the VdAL 1 Water content (%) 90.22 91.43 2 Ca (mg/L) 20.13 22.70 3 Ca content (mg/100 gFw) 9.83 9.71 4 Cu (mg/L) 0.13 0.15 5 Cu content (mg/100 gFw) 0.06 0.06 6 Fe (mg/L) 0.50 0.60 7 Fe content (mg/100 gFw) 2.50 2.98 8 K (mg/L) 158.00 184.00 9 K content (mg/100 gFw) 77.19 78.64 10 Mg (mg/L) 22.27 22.50 11 Mg content (mg/100 gFw) 10.88 9.63 12 Mn (mg/L) 0.10 0.12 13 Mn content (mg/100 gFw) 0.05 0.05 14 Na (mg/L) 7.20 7.35 15 Na content (mg/100 gFw) 3.52 3.14 16 P (mg/L) 18.30 20.13 17 P content (mg/100 gFw) 8.93 8.65 18 Zn (mg/L) 0.25 0.30 19 Zn content (mg/100 gFw) 0.12 0.13 Note: The content of each trace element in lines 3, 5, 7, 9, 11, 13, 15, 17 and 19 referred to the mass of each trace element of fresh watermelon per liter. The content of each trace element in lines 2, 4, 6, 8, 10, 12, 14, 16 and 18 referred to the content of each trace element in fresh watermelon.
(76) The results showed that the water content of watermelon was slightly increased by 1.34%, the Vc content was remarkably improved by 50.41%, the soluble sugar content was improved by 8.88%, the protein content of dry matters was improved by 14.2%, the protein content of fresh watermelon was improved by 0.62%, the titratable acid content and the soluble solid content remained unchanged, the dry matter content reduced by 12.4%, and the coarse fiber content reduced slightly after watermelon was treated with the agent of VdAL. It indicated that the contents of main nutrients in watermelon were increased, while the acidity was not significantly increased, and the dry matter content and the coarse fiber content decreased respectively, after watermelon was treated with the agent of VdAL, so that the palatability of watermelon was improved. After watermelon was treated with the agent of VdAL, the masses of Ca, Cu, Fe, K, Mg, Mn, Na, P and Zn in fresh watermelon per liter were increased by 12.75%, 16.54%, 19.21%, 16.46%, 1.05%, 20.45%, 2.13%, 10.02% and 19.73% respectively, and the mass contents of Cu, Fe, K, Mn and Zn in fresh watermelon were increased by 2.09%, 19.09%, 1.88%, 5.72% and 5.07% respectively. It indicated that the agent of VdAL could significantly improve the contents of trace elements Cu. Fe, K, Mn and Zn, which was beneficial to human body, in watermelon. The above experiment proved that the agent of VdAL could improve the quality of watermelon.
Example 4 the Agent of VdAL can Promote the Growth of Wheat and Improve the Yield of Wheat
(77) I. The Agent of VdAL can Promote the Germination and Vegetative Growth of Wheat
Experiment I
(78) The seeds of variety, Zhongmai 816, were respectively soaked into a 1000 times diluted VdAL, a 2000 times diluted VdAL, a 5000 times diluted VdA and a 10000 times diluted VdAL of Example 1 and clear water for 12 hours, and then accelerate germination for 96 hours at 25 C. in germination boxes, in which two layers of sterile filter paper wetted with sterile water were spread. The germination rate of the wheat seeds was statistically analyzed. The germination standard of the wheat seeds was that the bud length was equal to or more than half of the seed length, and root length was equal to or greater than seed length. 100 seeds were treated in each treatment. The experiment was repeated three times.
(79) The results showed that the germination rates of the wheat seeds of the contrast (i.e. the germination rates of the wheat seeds treated with clear water). VdAL of 1000 times dilution, VdAL of 2000 times dilution, VdAL of 5000 times dilution, and VdAL of 10000 times dilution were 95.311%, 83.261%, 92.511%, 97.671% and 99.5011% respectively. After the wheat seeds were treated with the 1000 times diluted VdAL, the 2000 times diluted VdAL, the 5000 times diluted VdA and the 10000 times diluted VdAL, the germination rates were improved by 12.64%, 0.03%, 2.48% and 4.40% respectively. It indicated that the agent of VdAL with proper concentration can promote germination of wheat seeds.
Experiment II
(80) The seeds of the variety of wheat, Zhongmai 816, were respectively soaked into a 5000 times diluted VdAL and a 10000 times diluted VdAL of Example 1 and clear water for 24 hours, and then were sowed into soil. The day of sowing was recorded as the first day. The wheat seedlings were taken out of the soil on the 3.sup.rd day after sowing. The fresh weight of the wheat seedlings was weighed.
(81) The result showed that the fresh weights of individual plant of wheat treated with the 5000 times diluted VdAL, the 10000 times diluted VdAL and clear water were 0.1280.022 g, 0.1410.022 g, and 0.1110.022 g respectively. After the seeds were soaked into the 5000 times diluted VdAL and the 10000 times diluted VdAL, the fresh weights of individual plant of wheat were improved by 15.32% and 27.03% respectively. The fresh weight of individual plant of wheat soaked into the 10000 times diluted VdAL was significantly higher than that of wheat soaked into clear water. It indicated that the agent of VdAL can promote vegetative growth of wheat.
(82) 2. The Agent of VdAL can Improve the Yield of Wheat
(83) Field experiment was designed as follows. The experiment adopted randomized block design. Three replicate blocks provided. Each replicate block was randomly provided with five treatment areas, which were respectively VdAL treatment area I, VdAL treatment area II, VdAL treatment area III, VdAL treatment area IV and contrast treatment area. The area of each treatment area was 0.5 mu.
(84) Wheat was treated once at the flowering stage of the seeds of the variety, Zhongmai 816, in VdAL treatment area I according to the following method. VdAL of 2500 times dilution of Example 1 with 45-50 kg per mu was sprayed to wheat leaves by the means of top spraying (mechanical spraying) to obtain wheat treated with a 2500 times diluted VdAL. The spraying was performed on morning or at evening, and preferably not at the high temperature period at noon. The agent of VdAL was preferably not mixed with pesticides and fertilizers at spraying. The agent of VdAL should be sprayed once again if it rained within 2 hours after being sprayed.
(85) According to the treatment method of VdAL treatment area I, the 2500 times diluted VdAL was replaced with a 5000 times diluted VdAL to treat VdAL treatment area II. Other steps remained unchanged. Wheat treated with the 5000 times diluted VdAL was obtained.
(86) According to the treatment method of VdAL treatment area I, the 2500 times diluted VdAL was replaced with a 10000 times diluted VdAL to treat VdAL treatment area III. Other steps remained unchanged. Wheat treated with the 10000 times diluted VdAL was obtained.
(87) According to the treatment method of VdAL treatment area I, the 2500 times diluted VdAL was replaced with a 20000 times diluted VdAL to treat VdAL treatment area IV. Other steps remained unchanged. Wheat treated with the 20000 times diluted VdAL was obtained.
(88) According to the treatment method of VdAL treatment area I, the 2500 times diluted VdAL was replaced with clear water to treat the contrast treatment area. Other steps remained unchanged. Untreated wheat was obtained.
(89) The yields of wheat in each treatment were statistically analyzed. The results showed that the yield of wheat treated with the 2500 times diluted VdAL was 8679 kg/ha, not improved; the average yields of wheat treated with the 5000 times diluted VdAL, the 10000 times diluted VdAL, the 20000 times diluted VdAL and clear water were 8942 kg/ha, 9485 kg/ha, 9231 kg/ha and 8772 kg/ha respectively; and after the wheat was treated with the 5000 times diluted VdAL, the 10000 times diluted VdAL, the 20000 times diluted VdAL, the yields were improved by 1.94%, 8.13% and 5.23% respectively.
(90) The thousand-grain weight, the number of effective spikes, the grain number per spike and the length of spike of wheat were statistically analyzed. The results were shown in Table 7. The result showed that, after the wheat was treated with the 5000 times diluted VdAL, the number of effective spikes was increased by 1.5%, and the grain number per spike was increased by 0.5%; after the wheat was treated with the 10000 times diluted VdAL, the number of effective spikes was increased by 7%, and the grain number per spike was increased by 3.5%; and after the wheat was treated with the 20000 times diluted VdAL, the number of effective spikes was increased by 2.3%, and the grain number per spike was increased by 3.7%.
(91) TABLE-US-00007 TABLE 7 the means of the thousand-grain weight, the number of effective spikes, the grain number per spike and the length of spike of wheat in different treatment Number of effective Thousand-grain spikes weight Number Grain number per spike Thousand- of effective Grain number Length of spike grain weight Increasing spikes(spikes/ Increasing per spike Increasing Length of Increasing Treatment (g) rate (%) ha) rate (%) (grains/pike) rate (%) spike (cm) rate (%) CK 37.69 / 1293.33 / 30.96 / 8.56 / Treatment 1 37.5 0.5 1322.67 2.3 32.12 3.7 8.53 0.3 Treatment 2 36.86 2.2 1384.33 7 32.05 3.5 8.65 1 Treatment 3 37.68 0 1313 1.5 31.13 0.5 8.58 0.2 Treatment 4 37.6 0.1 1289.33 0.4 30.74 0.7 8.78 2.6 Note: Treatment 1 represented the treatment with the 20000 times diluted VdAL, Treatment 2 represented the treatment with the 10000 times diluted VdAL, Treatment 3 represented the treatment with the 5000 times diluted VdAL, Treatment 4 represented the treatment with the 2500 times diluted VdAL, and CK represented untreatment (i.e., the treatment with clear water).
Example 5 the Agent of VdAL can Increase the Fruit Number of Individual Plant of Tomato
(92) Field experiment was designed as follows. The experiment adopted randomized block design. Three replicate blocks were provided. Each replicate block was randomly provided with three treatment areas, which were respectively VdAL treatment area I, VdAL treatment area II and contrast treatment area. The area of each treatment area was 0.1 mu.
(93) The VdAL was respectively sprayed to the variety of tomato, Fux, at early flowering stage and on the 15.sup.th day after the first treatment in VdAL treatment area I according to the following method. A 1000 times diluted VdAL of Example 1 with 45-50 kg per mu was sprayed to tomato leaves by the means of top spraying (mechanical spraying) to obtain tomato treated with the 1000 times diluted VdAL. The day of first spraying was recorded as 0 day after spraying. The spraying was performed on morning or at evening, and preferably not at the high temperature period at noon. The agent of VdAL was preferably not mixed with pesticides and fertilizers at spraying. The agent of VdAL should be sprayed once again if it rained within 2 hours after being sprayed.
(94) According to the treatment method of VdAL treatment area I, the 1000 times diluted VdAL was replaced with a 3000 times diluted VdAL to treat VdAL treatment area II. Other steps remained unchanged. Tomatoes treated with the 3000 times diluted VdAL were obtained.
(95) According to the treatment method of VdAL treatment area I, the 1000 times diluted VdAL was replaced with clear water to treat the contrast treatment area. Other steps remained unchanged. Untreated tomatoes were obtained.
(96) The fruit number of individual plant of tomato was statistically analyzed on the 20.sup.th day after spraying. The result showed that the average fruit number of individual plant of tomato treated with VdAL of 1000 times dilution, VdAL of 3000 times dilution and clearwater were 34.9, 40.4 and 33.3 respectively, and the fruit number of individual plant of tomato treated with VdAL of 1000 times dilution and VdAL of 3000 times dilution were improved by 4.8% and 21.3% respectively. This indicated that the agent of VdAL could promote tomato fruit.
Example 6 the Agent of VdAL can Promote the Growth of Green-Stem Cabbage
(97) Field experiment was designed as follows. The experiment adopted randomized block design. Three replicate blocks were provided. Each replicate block was randomly provided with five treatment areas, which were respectively VdAL treatment area I, VdAL treatment area II, VdAL treatment area III, VdAL treatment area IV and contrast treatment area. The area of each treatment area was 0.2 mu.
(98) The VdAL was sprayed once to the variety of green-stem cabbage, Huali, in VdAL treatment area I at two-leaf one-tip stage according to the following method. A 2000 times diluted VdAL of Example 1 with 45-50 kg per mu was sprayed to green-stem cabbage leaves by the means of top spraying (manual spraying) to obtain green-stem cabbage treated with the 2000 times diluted VdAL. The day of spraying the VdAL was recorded as 0 day of treatment. The spraying was performed on morning or at evening, and preferably not at the high temperature period at noon. The agent of VdAL was preferably not mixed with pesticides and fertilizers at spraying. The agent of VdAL should be sprayed once again if it rained within 2 hours after being sprayed.
(99) According to the treatment method of VdAL treatment area I, the 2000 times diluted VdAL was replaced with a 2500 times diluted VdAL to treat VdAL treatment area II. Other steps remained unchanged. Green-stem cabbages treated with the 2500 times diluted VdAL were obtained.
(100) According to the treatment method of VdAL treatment area I, the 2000 times diluted VdAL was replaced with a 3000 times diluted VdAL to treat VdAL treatment area III. Other steps remained unchanged. Green-stem cabbages treated with the 3000 times diluted VdAL were obtained.
(101) According to the treatment method of VdAL treatment area I, the 2000 times diluted VdAL was replaced with a 4000 times diluted VdAL to treat VdAL treatment area IV. Other steps remained unchanged. Green-stem cabbages treated with the 4000 times diluted VdAL were obtained.
(102) According to the treatment method in VdAL, treatment area I, the 2000 times diluted VdAL was replaced with clear water to treat the contrast treatment area. Other steps remained unchanged. Untreated green-stem cabbages were obtained.
(103) The fresh weights of 50 green-stem cabbages in different treatments were statistically analyzed on the 20.sup.th day after the treatment. The result showed that the average fresh weights of 50 green-stem cabbages treated with the 2000 times diluted VdAL, the 2500 times diluted VdAL, the 3000 times diluted VdAL, the 4000 times diluted VdAL and clear water were 2.960 kg, 3.270 kg, 3.835 kg, 3.895 kg and 1.912 kg respectively; and the fresh weights of 50 green-stem cabbages treated with the 2000 times diluted VdAL, the 2500 times diluted VdAL, the 3000 times diluted VdAL, and the 4000 times diluted VdAL were improved by 54.8%, 71.0%, 100.6% and 103.7% respectively. This indicated that the agent of VdAL could promote the growth of green-stem cabbage.
Example 7 Effects of the Agent of VdAL in Other Crops
(104) 1. The Agent of VdAL can Promote the Growth of Cucumber, Improve the Yield and Delay Aging Thereof
(105) Field experiment was designed as follows. The experiment adopted randomized block design. Three replicate blocks were provided. Each replicate block was randomly provided with five treatment areas, which were respectively VdAL treatment area I, VdAL treatment area II, VdAL treatment area III, VdAL treatment area IV and contrast treatment area. The area of each treatment area was 0.1 mu.
(106) The VdAL was sprayed to the variety of cucumber, Jinza No. 1 in VdAL treatment area I at early flowering stage and on the 15.sup.th day after the first treatment respectively according to the following method. A 300 times diluted VdAL of Example 1 with 45-50 kg per mu was sprayed to cucumber leaves by the means of top spraying (manual spraying) to obtain cucumber treated with the 300 times diluted VdAL. The day of the first time of spraying VdAL was recorded as 0 day after spraying. The spraying was performed on morning or at evening, and preferably not at the high temperature period at noon. The agent of VdAL was preferably not mixed with pesticides and fertilizers at spraying. The agent of VdAL should be sprayed once again if it rained within 2 hours after being sprayed.
(107) According to the treatment method of VdAL treatment area I, the 300 times diluted VdAL of was replaced with a 600 times diluted VdAL to treat VdAL treatment area II. Other steps remained unchanged. Cucumbers treated with the 600 times diluted VdAL were obtained.
(108) According to the treatment method of VdAL treatment area I, the 300 times diluted VdAL of was replaced with a 900 times diluted VdAL to treat VdAL treatment area III. Other steps remained unchanged. Cucumbers treated with the 900 times diluted VdAL were obtained.
(109) According to the treatment method of VdAL treatment area I, the 300 times diluted VdAL of was replaced with a 2000 times diluted VdAL to treat VdAL treatment area IV. Other steps remained unchanged. Cucumbers treated with the 2000 times diluted VdAL were obtained.
(110) According to the treatment method of VdAL treatment area I, the 300 times diluted VdAL was replaced with clear water to treat the contrast treatment area. Other steps remained unchanged. Untreated cucumbers were obtained.
(111) The result of cucumber was statistically analyzed on the 20.sup.th day after spraying. The result showed that the agent of VdAL could improve the fruit setting rate of cucumber, promote the expansion of cucumber and improve the yield of cucumber (
(112) 2. The Agent of VdAL can Improve the Fruit Setting Rate and Yield of Cherry Tomato
(113) Field experiment was designed as follows. The experiment adopted randomized block design. Three replicate blocks were provided. Each replicate block was randomly provided with two treatment areas, which were respectively VdAL treatment area and contrast treatment area. The area of each treatment area was 1 mu.
(114) The VdAL was sprayed once to the variety of cherry tomato, Jingdan No. 2, at flowering stage in VdAL treatment area according to the following method. A 2000 times diluted VdAL of Example 1 with 45-50 kg per mu was sprayed to cherry tomato leaves by the means of top spraying (manual spraying) to obtain cherry tomato treated with the 2000 times diluted VdAL. The day of spraying the agent of VdAL was recorded as 0 day of the treatment. The spraying was performed on morning or at evening, and preferably not at the high temperature period at noon. The agent of VdAL was preferably not mixed with pesticides and fertilizers at spraying. The agent of VdAL should be sprayed once again if it rained within 2 hours after being sprayed.
(115) According to the treatment method of VdAL treatment area, the 2000 times diluted VdAL was replaced with clear water to treat the contrast treatment area. Other steps remained unchanged. Untreated cherry tomatoes were obtained.
(116) The result of cherry tomato was observed on the 7-10.sup.th day after treatment. The result showed that the agent of VdAL could improve the fruit setting rate and yield of cherry tomato (
(117) 3. The Agent of VdAL can Promote the Growth of Strawberry
(118) Field experiment was designed as follows. The variety of strawberry was Fengxiang. The experiment adopted randomized block design. Three replicate blocks were provided. Each replicate block was randomly provided with two treatment areas, which were respectively VdAL treatment area and contrast treatment area. The area of each treatment area was 1 mu.
(119) The VdAL was sprayed once to the variety of strawberry, Fengxiang, at flowering stage in VdAL treatment area according to the following method. A 2000 times diluted VdAL of Example 1 with 45-50 kg per mu was sprayed to strawberry leaves by the means of top spraying (manual spraying) to obtain strawberry treated with the 2000 times diluted VdAL. The day of spraying the agent of VdAL was recorded as 0 day of the treatment. The spraying was performed on morning or at evening, and preferably not at the high temperature period at noon. The agent of VdAL was prohibited to be mixed with pesticides and fertilizers at spraying. The agent of VdAL should be sprayed once again if it rained within 2 hours after being sprayed.
(120) According to the treatment method of VdAL treatment area, the 2000 times diluted VdAL was replaced with clear water to treat the contrast treatment area. Other steps remained unchanged. Untreated strawberries were obtained.
(121) The growth situation of strawberry was observed on the 7-10.sup.th day after treatment. The result showed that the agent of VdAL could promote vegetative growth of strawberry (
(122) 4. The Agent of VdAL can Promote the Growth of Melon
(123) Field experiment was designed as follows. The variety of melon was Super Cuibaoxiang. The experiment adopted randomized block design. Three replicate blocks were provided. Each replicate block was randomly provided with two treatment areas, which were respectively VdAL treatment area and contrast treatment area. The area of each treatment area was 1 mu.
(124) The VdAL was sprayed once to the variety of melon, Super Cuibaoxiang, at seedling stage in VdAL treatment area according to the following method. A 2000 times diluted VdAL of Example 1 with 45-50 kg per mu was sprayed to melon leaves by the means of top spraying (manual spraying) to obtain melon treated with the 2000 times diluted VdAL. The day of spraying the agent of VdAL was recorded as 0 day of the treatment. The spraying was performed on morning or at evening, and preferably not at the high temperature period at noon. The agent of VdAL was prohibited to be mixed with pesticides and fertilizers at spraying. The agent of VdAL should be sprayed once again if it rained within 2 hours after being sprayed.
(125) According to the treatment method of VdAL treatment area, the 2000 times diluted VdAL was replaced with clear water to treat the contrast treatment area. Other steps remained unchanged. Untreated melons were obtained.
(126) The growth situation of melon was observed on the 30.sup.th day after treatment. The result showed that VdAL could promote vegetative growth of melon (
(127) 5. The Agent of VdAL can Promote the Growth of Sweet Pepper
(128) Field experiment was designed as follows. The experiment adopted randomized block design. Three replicate blocks were provided. Each replicate block was randomly provided with two treatment areas, which were respectively VdAL treatment area and contrast treatment area. The area of each treatment area was 1 mu.
(129) The VdAL was sprayed once to the variety of sweet pepper, Ruiyou 816, at flowering stage in VdAL treatment area according to the following method. A 1000 times diluted VdAL of Example 1 with 45-50 kg per mu was sprayed to sweet pepper leaves by the means of top spraying (manual spraying) to obtain sweet pepper treated with the 1000 times diluted VdAL. The day of spraying the agent of VdAL was recorded as 0 day of the treatment. The spraying was performed on morning or at evening, and preferably not at the high temperature period at noon. The agent of VdAL was prohibited to be mixed with pesticides and fertilizers at spraying. The agent of VdAL should be sprayed once again if it rained within 2 hours after being sprayed.
(130) According to the treatment method of VdAL treatment area, the 1000 times diluted VdAL was replaced with clear water to treat the contrast treatment area. Other steps remained unchanged. Untreated sweet peppers were obtained.
(131) The growth situation of sweet pepper was observed on the 7-10.sup.th day after treatment. The result showed that the sweet pepper was vigorous in growth and its leaves were dark green after the agent of VdAL was sprayed (
(132) 6. The Agent of VdAL can Promote Seedling Emergence and Growth of Soybean
(133) The seeds of the variety of soybean, King of China soybean, were soaked into a 1000 times diluted VdAL, a 3000 times diluted VdAL, a 5000 times diluted VdAL, a 10000 times diluted VdAL and a 20000 times diluted VdAL of Example 1 and clear water, respectively, for 24 hours, and then accelerate germination for 72 hours at 25 C. in trays, in which two layers of sterile filter paper wetted with sterile water were spread. The germination rate of the soybean seeds was statistically analyzed. 100 seeds were treated in each treatment. The experiment was repeated three times.
(134) The result showed (
Experiment II
(135) The seeds of the variety of soybean, King of China soybean, were soaked into a 5000 times diluted VdAL, of Example 1 and clear water, respectively, for 24 hours, and then were sowed into soil. The day of sowing was recorded as the first day after sowing. Soybean seedlings were taken out of the soil on the 20.sup.th day after sowing. The fresh weight of the soybean seedlings was weighed (
(136) The result showed that the fresh weights of individual plant of soybean treated with the 5000 times diluted VdAL and clear water were 1.590.3 g and 1.360.3 g respectively; after the seeds were soaked into the 5000 times diluted VdAL, the fresh weight of individual plant of soybean was improved by 16.91%; the fresh weights of the ground part of individual plant of soybean treated with the 5000 times diluted VdAL and clear water were 1.360.3 g and 1.060.3 g respectively; and after the seeds were soaked into the 5000 times diluted VdAL, the fresh weight of the ground part of individual plant of soybean was improved by 28.30%, which was significantly higher than the fresh weight of the ground part of individual plant of soybean treated with clear water. This indicated that the agent of VdAL could promote vegetative growth of soybean.
Example 8 the Agent of VdAL can Promote the Growth of Rice and Improve the Yield Thereof
(137) Field experiment was designed as follows. The experiment adopted randomized block design. Three replicate blocks were provided. Each replicate block was randomly provided with four treatment areas, which were respectively VdAL treatment area I, VdAL treatment area II, VdAL treatment area III and contrast treatment area. The area of each treatment area was 0.1 mu.
(138) The variety of rice, Nongda 502, of VdAL treatment area I was treated once at flowering stage according to the following method. A 10000 times diluted VdAL of Example 1 with 45-50 kg per mu was sprayed to rice leaves by the means of top spraying (manual spraying) to obtain rice treated with the 10000 times diluted VdAL (1/10000). The spraying was performed on morning or at evening, and preferably not at the high temperature period at noon. The agent of VdAL was prohibited to be mixed with pesticides and fertilizers at spraying. The agent of VdAL should be sprayed once again if it rained within 2 hours after being sprayed.
(139) According to the treatment method of VdAL treatment area I, the 10000 times diluted VdAL was replaced with a 20000 times diluted VdAL to treat VdAL treatment area II. Other steps remained unchanged. Rice treated with the 20000 times diluted VdAL (1/20000) was obtained.
(140) According to the treatment method of VdAL treatment area I, the 10000 times diluted VdAL was replaced with a 40000 times diluted VdAL to treat VdAL treatment area III. Other steps remained unchanged. Rice treated with the 40000 times diluted VdAL (1/40000) was obtained.
(141) According to the treatment method of VdAL treatment area I, the 10000 times diluted VdAL was replaced with clear water to treat the contrast treatment area. Other steps remained unchanged. Untreated rice (CK) was obtained.
(142) The yields of rice in each treatment were statistically analyzed. The result (
Example 9 the Agent of VdAL can Improve the Drought Resistance of Wheat
(143) The experiments were repeated three times. The specific steps of each repeated experiment were shown as follows.
Experiment I
(144) The seeds of wheat, Zhongmai 816, were divided into four groups, i.e. group 1, group 2, group 3 and group 4. Each group had 50 seeds.
(145) The agent of VdAL of Example 1 was added into Fuwu (Beinong (Haili) Zhuozhou Seed Coat Co., Ltd.) having a mass percentage concentration of 6% to obtain the liquid in which the mass percentage concentration of the agent of VdAL was 2%. The liquid was named 2.0%+Fuwu. Wheat seeds of group 1 were treated by 2.0%+Fuwu according to the following method. 100-200 g of 2.0%+Fuwu was blended with 1.5-2 kg of water, and was stirred uniformly, then was mixed with 100 kg of seeds, and the following was to be stirred uniformly. The seeds were dried in the shade for sowing. Then the wheat seeds of group 1 were sowed into soil. Watering wheat was stopped (i.e., water-control treatment) at the three-leaf stage of wheat.
(146) The agent of VdAL of Example 1 was added into Fuwu having a mass percentage concentration of 6% to obtain the liquid in which the mass percentage concentration of the agent of VdAL was 1%. The liquid was named 1.0%+Fuwu. Wheat seeds of group 2 were treated by 1.0%+Fuwu according to the following method. 100-200 g of 1.0%+Fuwu was blended with 1.5-2 kg of water, and was stirred uniformly, then was mixed with 100 kg of seeds, and the following was to be stirred uniformly. The seeds were dried in the shade for sowing. Then the wheat seeds of group 2 were sowed into soil. Watering wheat was stopped (i.e., water-control treatment) at the three-leaf stage of wheat.
(147) Wheat seeds of group 3 were treated by 1.0%+Fuwu according to the following method. 100-200 g of Fuwu (Beinong (Haili) Zhuozhou Seed Coat Co., Ltd.) having a mass percentage concentration of 6% was blended with 1.5-2 kg of water, and was stirred uniformly, then was mixed with 100 kg of seeds, and the following was to be stirred uniformly. The seeds were dried in the shade for sowing. Then the wheat seeds of group 3 were sowed into soil. Watering wheat was stopped (i.e., water-control treatment) at the three-leaf stage of wheat.
(148) Wheat seeds of group 4 were treated with clear water as a contrast (CK) according to the following method. 100 kg of seeds were mixed with 1.5-2 kg of water, and were stirred uniformly. The seeds were dried in the shade for sowing. Then the wheat seeds of group 4 were sowed into soil. Watering wheat was stopped (i.e., water-control treatment) at the three-leaf stage of wheat.
(149) The first day of stopping watering was recorded as 1.sup.st day of water-control. The result showed that the agent VdAL added into a seed coating to treat wheat seeds could significantly improve drought resistance of wheat. All of the wheat of group 1 died within 20 days after water-control. All of the wheat of group 2 died within 15 days after water-control. All of the wheat of group 3 died within 10 days after water-control. All of the wheat of group 4 died within 12 days after water-control. The time when all of the wheat of group 1 died was significantly longer than the time when all of the wheat of group 3 and group 4 died, respectively. The time when all of the wheat of group 2 died was significantly longer than the time when all of the wheat of group 3 and group 4 died, respectively. The growth situation of wheat of each group on 7 days after water-control was shown in
Experiment II
(150) The seeds of wheat, Zhongmai 816, were divided into six groups, i.e. group 1, group 2, group 3, group 4, group 5 and group 6. Each group had 50 seeds.
(151) Wheat seeds of group 1 were treated with clear water as a contrast (CK1) according to the following method. 100 kg of seeds were mixed with 1.5-2 kg of clear water, and were stirred uniformly. The seeds were dried in the shade for sowing. Then the wheat seeds of group 1 were sowed into soil. Watering wheat was stopped at three-leaf stage. The wheat was re-watered on the 7.sup.th day after watering was stopped.
(152) The agent of VdAL of Example 1 was added into Fuwu having a mass percentage concentration of 6% to obtain the liquid in which the mass percentage concentration of the agent of VdAL was 0.2%. The liquid was named 0.2%+Fuwu. Wheat seeds of group 2 were treated by 0.2%+Fuwu according to the following method. 100-200 g of 0.2%+Fuwu was blended with 1.5-2 kg of water, and was stirred uniformly, then was mixed with 100 kg of seeds, and the following was to be stirred uniformly. The seeds were dried in the shade for sowing. Then the wheat seeds of group 2 were sowed into soil. Watering wheat was stopped at three-leaf stage. The wheat was re-watered on the 7.sup.th day after watering was stopped.
(153) The VdAL of Example 1 was added into Fuwu having a mass percentage concentration of 6% to obtain the liquid in which the mass percentage concentration of the agent of VdAL was 1%. The liquid was named 1.0%+Fuwu. Wheat seeds of group 3 were treated by 1.0%+Fuwu according to the following method. 100-200 g of 1.0%+Fuwu was blended with 1.5-2 kg of water, and was stirred uniformly, then was mixed with 100 kg of seeds, and the following was to be stirred uniformly. The seeds were dried in the shade for sowing. Then the wheat seeds of group 3 were sowed into soil. Watering wheat was stopped at three-leaf stage. The wheat was re-watered on the 7.sup.th day after watering was stopped.
(154) The VdAL of Example 1 was added into Fuwu having a mass percentage concentration of 6% to obtain the liquid in which the mass percentage concentration of the agent of VdAL was 2%. The liquid was named 2.0%+Fuwu. Wheat seeds of group 4 were treated by 2.0%+Fuwu according to the following method. 100-200 g of 2.0%+Fuwu was blended with 1.5-2 kg of water, and was stirred uniformly, then was mixed with 100 kg of seeds, and the following was to be stirred uniformly. The seeds were dried in the shade for sowing. Then the wheat seeds of group 4 were sowed into soil. Watering wheat was stopped at three-leaf stage. The wheat was re-watered on the 7.sup.th day after watering was stopped.
(155) The VdAL of Example 1 was added into Fuwu having a mass percentage concentration of 6% to obtain the liquid in which the mass percentage concentration of the agent of VdAL was 4%. The liquid was named 4.0%+Fuwu. Wheat seeds of group 5 were treated by 4.0%+Fuwu according to the following method. 100-200 g of 4.0%+Fuwu was blended with 1.5-2 kg of water, and was stirred uniformly, then was mixed with 100 kg of seeds, and the following was to be stirred uniformly. The seeds were dried in the shade for sowing. Then the wheat seeds of group 5 were sowed into soil. Watering wheat was stopped at three-leaf stage. The wheat was re-watered on the 7.sup.th day after watering was stopped.
(156) Wheat seeds of group 6 were treated by 6% Fuwu as a contrast (CK2) according to the following method. 100-200 g of Fuwu (Beinong (Haili) Zhuozhou Seed Coat Co., Ltd.) having a mass percentage concentration of 6% was blended with 1.5-2 kg of water, and was stirred uniformly, then was mixed with 100 kg of seeds, and the following was to be stirred uniformly. The seeds were dried in the shade for sowing. Then the wheat seeds of group 6 were sowed into soil. Watering wheat was stopped at three-leaf stage. The wheat was re-watered on the 7.sup.th day after watering was stopped.
(157) The chlorophyll content of wheat leaves of each group was determined by SPAD-502Plus (Beijing Bolun Jingwei Science and Technology Development Co., Ltd.) on the same day that watering for each group was stopped. The result was shown in
(158) The re-watering index was calculated on the 4.sup.th day after re-watering treatment for each group, wherein the re-watering index=(1I-level plant number+2II-level plant number+3III-level plant number+4IV-level plant number)/(4investigated plant number).
(159) The grading standards of plant were as follows:
(160) Level 0: after watering was recovered, the curling area of individual leaf was 100%, and leaf did not become green;
(161) Level I: after watering was recovered, the curling area of individual leaf was 100%, and leaf started to become green;
(162) Level II: after watering was recovered, the curling area of individual leaf was more than or equal to 50% and less than 100%;
(163) Level III: after watering was recovered, the curling area of individual leaf was more than or equal to 0% and less than 50%;
(164) Level IV: after watering was recovered, the curling area of individual leaf was 0%.
(165) The result was as shown
Example 10 the Agent of VdAL can Improve the Quality of Grape
(166) Field experiment was designed as follows. The place for experiment was Dunhuang, Gansu province. The experiment adopted randomized block design. Three replicate blocks were provided. Each replicate block was randomly provided with two treatment areas, which were respectively VdAL treatment area and contrast treatment area. The area of each treatment area was 0.1 mu.
(167) The variety of grape red globe (Gansu Yongdeng Yongsheng Red Grape Professional Cooperative) was treated once at flowering stage in VdAL treatment area according to the following method. A 3000 times diluted VdAL of Example 1 with 45-50 kg per mu was sprayed to grape leaves by the means of top spraying (manual spraying) to obtain grape treated with the 3000 times diluted VdAL (1/3000). The spraying was performed on morning or at evening, and preferably not at the high temperature period at noon. The agent of VdAL was prohibited to be mixed with pesticides and fertilizers at spraying. The agent of VdAL should be sprayed once again if it rained within 2 hours after being sprayed.
(168) According to the treatment method of VdAL treatment area, the 3000 times diluted VdAL was replaced with clear water to treat the contrast treatment area. Other steps remained unchanged. Untreated grapes were obtained (contrast, CK).
(169) The mature time of grape in each treatment was statistically analyzed. The result showed that the average mature times of grape treated with the 3000 times diluted VdAL and clear water were September 15 (85 days after flowering stage, or 75 days after VdAL treatment) and September 20 (90 days after flowering stage, or 80 days after clear water treatment), respectively. The mature time of grape treated with the 3000 times diluted VdAL was 5 days early.
(170) The Vc content of the mature grape was determined according to a 2,6-dichlorophenol indophenol titration method. The result showed that the average Vc contents of grape treated with the 3000 times diluted VdAL and clear water were respectively 18.20 mg/100 g fresh weight and 12.10 mg/100 g fresh weight. The Vc content of the grape treated with the 3000 times diluted VdAL was improved by 50.4%.
(171) By observing mature grape, it showed that the color of grape sprayed VdAL was brighter and the bunch of grape was compact and the fruit dropped down uneasily (
Example 11 the Agent of VdAL can Promote the Maturity and Coloration of Cherry and Improve the Yield of Cherry
(172) Field experiment was designed as follows. The place of experiment was Fusban District, Yantai City. The experiment adopted randomized block design. Three replicate blocks were provided. Each replicate block was randomly provided with two treatment areas, which were respectively VdAL treatment area and contrast treatment area. The area of each treatment area was 0.1 mu.
(173) The variety of cherry, Longguan (Yantai Qixia Qunli Fruits and Nursery Stock Professional Cooperatives), was treated once at flowering stage in VdAL treatment area according to the following method. A 5000 times diluted VdAL of Example 1 with 45-50 kg per mu was sprayed to cherry leaves by the means of top spraying (manual spraying) to obtain cherries treated with the 5000 times diluted VdAL (1/5000). The spraying was performed on morning or at evening, and preferably not at the high temperature period at noon. The agent of VdAL was prohibited to be mixed with pesticides and fertilizers at spraying. The agent of VdAL should be sprayed once again if it rained within 2 hours after being sprayed.
(174) According to the treatment method of VdAL treatment area, the 5000 times diluted VdAL was replaced with clear water to treat the contrast treatment area. Other steps remained unchanged. Untreated cherries were obtained (contrast, CK).
(175) The mature time of cherry in each treatment was statistically analyzed. The result showed that the average mature time of cherry treated with the 5000 times diluted VdAL and clear water was respectively May 1 (52 days after flowering stage and VdAL treatment) and May 6 (57 days after VdAL treatment). The mature time of cherry treated with the 5000 times diluted VdAL was 5 days early.
(176) The yield of cherry in each treatment was statistically analyzed. The result showed that the average yields of cherry treated with the 5000 times diluted VdAL and clear water were 2200 kg and 1715 kg respectively. The yield of cherry treated with the 5000 times diluted VdAL was improved by 28.28%.
(177) When cherry was mature (52 days after VdAL treatment, or 57 days after clear water treatment), the appearance of cherry was observed. The result showed that the color of the cherry treated with VdAL was significantly redder than that of untreated cherry (
Example 12 the Agent of VdAL can Promote the Growth of Can-Like Radish and Improve the Palatability Thereof
(178) Field experiment was designed as follows. The experiment adopted randomized block design. Three replicate blocks were provided. Each replicate block was randomly provided with two treatment areas, which were respectively VdAL treatment area and contrast treatment area. The area of each treatment area was 0.1 mu.
(179) The variety of can-like radish, 791 (Zhengzhou Sanhe Agricultural Science and Technology Development Co., Ltd.), was treated once at root expansion stage in VdAL treatment area according to the following method. A 5000 times diluted VdAL of Example 1 with 45-50 kg per mu was sprayed to can-like radish leaves by the means of top spraying (manual spraying) to obtain can-like radish treated with the 5000 times diluted VdAL (1/5000). The spraying was performed on morning or at evening, and preferably not at the high temperature period at noon. The agent of VdAL was prohibited to be mixed with pesticides and fertilizers at spraying. The agent of VdAL should be sprayed once again if it rained within 2 hours after being sprayed.
(180) According to the treatment method of VdAL treatment area, the 5000 times diluted VdAL was replaced with clear water to treat the contrast treatment area. Other steps remained unchanged. Untreated can-like radishes were obtained (contrast, CK).
(181) The yield of can-like radish in each treatment was statistically analyzed on the 20.sup.th day after VdAL/clear water treatment (
(182) 30 experienced evaluators were selected to evaluate the brittleness of the can-like radish treated with the above two methods (
(183) Other 30 experienced evaluators were selected to evaluate the sweetness of the can-like radish treated with the above two method. Apple red Fuji with a score of 10 for the brittleness was used as a reference sample. 10 parts of samples of each kind of can-like radish were selected for 10 parallel experiments. The mean of scores was calculated. The result showed that the average score of the sweetness of untreated can-like radish was 4.27, and the average score of the sweetness of can-like radish treated with the 5000 times diluted VdAL was 5.56. The average relative deviations of 10 parallel experiments were less than 20% after determination, respectively, which meet the experimental requirement. The average score of the sweetness of untreated can-like radish was significantly less than that of the can-like radish treated with the 5000 times diluted VdAL.
(184) This indicated that the agent of VdAL could improve the brittleness and sweetness of can-like radish and the can-like radish treated with the agent of VdAL had strong palatability.
Example 13 the Agent of VdAL can Promote the Growth of Spinach and Improve the Yield Thereof
(185) Field experiment was designed as follows. The experiment adopted randomized block design. Three replicate blocks were provided. Each replicate block was randomly provided with three treatment areas, which were respectively VdAL treatment area I, VdAL treatment area II and contrast treatment area. The area of each treatment area was 0.1 mu.
(186) The variety of spinach, Qiulv (Zhengzhou Sanhe Agricultural Science and Technology Development Co., Ltd.), was treated once at three-leaf stage in VdAL treatment area I according to the following method. A 4000 times diluted VdAL of Example 1 with 15-20 kg per mu was sprayed to spinach leaves by the means of top spraying (manual spraying) to obtain spinach treated with the 4000 times diluted VdAL (1/4000). The spraying was performed on morning or at evening, and preferably not at the high temperature period at noon. The agent of VdAL was prohibited to be mixed with pesticides and fertilizers at spraying. The agent of VdAL should be sprayed once again if it rained within 2 hours after being sprayed.
(187) According to the treatment method of VdAL treatment area I, the 4000 times diluted VdAL was replaced with a 6000 times diluted VdAL to treat VdAL treatment area II. Other steps remained unchanged. Spinaches treated with the 6000 times diluted VdAL (1/6000) were obtained.
(188) According to the treatment method of VdAL treatment area I, the 4000 times diluted VdAL was replaced with clear water to treat the contrast treatment area. Other steps remained unchanged. Untreated spinaches (CK) were obtained.
(189) The yield of spinach in each treatment was statistically analyzed on the 20.sup.th days after VdAL/clear water treatment. The result showed that the average yields of the spinach treated with the 4000 times diluted VdAL, the 6000 times diluted VdAL and clear water were 0.58 g/0.12 m.sup.2, 0.98 kg/0.12 m.sup.2 and 0.63 kg/0.12 m.sup.2 respectively, and the yields of the spinach treated with the 4000 times diluted VdAL and the 6000 times diluted VdAL were improved by 7.45% and 56.38% respectively, which indicated that VdAL with a proper concentration for treating spinach could improve the yield thereof.
INDUSTRIAL APPLICATION
(190) Experiments prove that the biological agent of VdAL according to the present invention, wherein VdAL is an active ingredient, can promote the seed germination, plant growth and fruit growth of multiple plant, improve the fruit setting number of multiple plant, and delay aging of multiple plant.
(191) 1. The average germination rate of cotton after cotton seeds are treated with the 7500 times diluted VdAL is 1.08 times that of cotton after cotton seeds are treated with clear water. When Lu No. 30 is treated with the 3000 times diluted VdAL, the cotton yield before first frost is improved by 19.48% and the total yield is improved by 17.04%. When the cotton was treated with the 5000 times diluted VdAL, the cotton yield of Lu No. 30 before first frost is improved by 21.25% and the total yield thereof is improved by 20.00%, the cotton yield of 616 before first frost is improved by 19.06% and the total yield thereof is improved by 19.69%, and the cotton yield of Lu No. 25 before first frost is improved by 16.37% and the total yield thereof is improved by 17.60%. When the cotton was treated with the 3000 times diluted VdAL, the number of boll setting of individual plant, total number of the boll per mu, ginning outturn and the yield per mu were increased by 4.5%, 1.98%, 0.24% and 1.76% respectively. When the cotton was treated with the 5000 times diluted VdAL, the number of boll setting of individual plant, total number of the boll per mu, the weight of single boll, ginning outturn and the yield per mu were increased by 3.41%, 4.41%, 1.9%, 0.49% and 5.22%, respectively.
(192) 2. After watermelon is treated with the 2000 times diluted VdLA, the fruit number of individual plant of watermelon is improved by 18.2%, and the yield is improved by 33.4%.
(193) 3. After wheat seeds are treated with the 1000 times diluted VdAL, the 2000 times diluted VdAL, the 5000 times diluted VdAL and the 10000 times diluted VdAL, the germination rates are improved by 2 times, 9 times, 12 times and 11 times, respectively. The germination rates of the wheat seeds treated with the 5000 times diluted VdAL and the 10000 times diluted VdAL are improved by 2.48% and 4.40% respectively. After seeds are soaked into the 5000 times diluted VdAL and the 10000 times diluted VdAL, the fresh weights of individual plant of wheat were improved by 15.32% and 27.03%, respectively. After wheat is treated with the 5000 times diluted VdAL, the 10000 times diluted VdAL and the 20000 times diluted VdAL, the yields are improved by 1.94%, 8.13% and 5.23%, respectively. After wheat is treated with the 5000 times diluted VdAL, the number of effective spike is increased by 1.5%, and the grain number per spike is increased by 0.5%; after the wheat is treated with the 10000 times diluted VdAL, the number of effective spike is increased by 7%, and the grain number per spike is increased by 3.5%; and after the wheat is treated with the 20000 times diluted VdAL, the number of effective spike is increased by 2.3%, and the grain number per spike is increased by 3.7%.
(194) 4. After tomatoes are treated with the 1000 times diluted VdAL and the 3000 times diluted VdAL, the fruit numbers of individual plant of tomato is improved by 4.8% and 21.3%, respectively.
(195) 5. After green-stem cabbage is treated with the 2000 times diluted VdAL, the 2500 times diluted VdAL, the 3000 times diluted VdAL and the 4000 times diluted VdAL, the fresh weights of 50 plant of green-stem cabbages are improved by 54.8%, 71.0%, 100.6% and 103.7%, respectively.
(196) 6. The VdAL can further improve the fruit setting rate of cucumber, promote the expansion of cucumber, improve the yield of cucumber, promote the growth of cucumber plant and delay aging of cucumber plant. The VdAL can further improve the fruit setting rate of cherry tomato, promote plant growth of strawberry, melon and sweet pepper, and promote germination and growth of soybean. After seeds are soaked into the 5000 times diluted VdAL, the fresh weight of individual plant of soybean is improved by 16.91%, and the ground part fresh weight of individual plant of soybean is improved by 28.30%.
(197) 7. After rice is treated with the 10000 times diluted VdAL and the 20000 times diluted VdAL, the yields are improved by 12.28% and 2.51%, respectively.
(198) 8. The average weights of individual root tuber of can-like radish treated with the 5000 times diluted VdAL and clear water are 0.90 kg and 0.85 kg, respectively. The average weight of individual root tuber of the can-like radish treated with the 5000 times diluted VdAL is improved by 5.88%. The average fresh weights of individual plant of can-like radish treated with the 5000-times diluted VdAL and clear water are 1.20 kg and 1.09 kg, respectively. The average fresh weight of individual plant of can-like radish treated with the 5000 times diluted VdAL is improved by 10.09%.
(199) 9. The average yields of spinach treated with the 4000 times diluted VdAL, the 6000 times diluted VdAL and clear water are 0.58 kg/0.12 m.sup.2, 0.98 kg/0.12 m.sup.2 and 0.63 kg/0.12 m.sup.2 respectively. The yields of the spinach treated with the 4000 times diluted VdAL and the 6000 times diluted VdAL are improved by 7.45% and 56.38%, respectively, which indicated that spinach treated with VdAL with a proper concentration can improve yield thereof.
(200) Experiments prove that the biological agent of VdAL according to the present invention, wherein VdAL is an active ingredient, can improve the quality of plant product and promote the fruit maturity.
(201) 1. After VdAL treatment, the contents of main nutrients and trace elements in watermelon are increased, and the palatability is improved. After VdAL treatment, the Vc content is remarkably improved by 50.41%; the soluble sugar content is improved by 8.88%; the protein content of dry matters is improved by 14.2%, the protein content of fresh watermelon is improved by 0.62%, the dry matter content declines by 12.4%, and the coarse fiber content declines a little. After VdAL treatment, the masses of Ca, Cu. Fe, K, Mg, Mn, Na, P and Zn in each liter of fresh watermelon are increased by 12.75%, 16.54%, 19.21%, 16.46%, 1.05%, 20.45%, 2.13%, 10.02% and 19.73%, respectively; and the mass contents of Cu, Fe, K, Mn and Zn in fresh watermelon are increased by 2.09%, 19.09%, 1.88%, 5.72% and 5.07%, respectively.
(202) 2. After VdAL treatment, the cotton yield before first frost and the ginning outturn are increased. When Lu No. 30 is treated with the 3000 times diluted VdAL, the cotton yield before first frost is improved by 19.48%; when being treated with the 5000 times diluted VdAL, the cotton yield of Lu No. 30 before first frost is improved by 21.25%, the cotton yield of 616 before first frost is improved by 19.06%, and the cotton yield of Lu No. 25 before first frost is improved by 16.37%. When being treated with the 3000 times diluted VdAL, the ginning outturn is increased by 0.24%; and when being treated with the 5000 times diluted VdAL, the ginning outturn is increased by 0.49%.
(203) 3. After grape is treated with the 3000 times diluted VdAL, the Vc content of the grape is improved by 50.4%, the grape is brighter in color, grape clusters are compact and unlikely to drop and the mature time of the grape is 5 days ahead of time.
(204) 4. After cherries are treated with the 5000 times diluted VdAL, the mature time of the cherries is 5 days ahead of time, and the yield is improved by 28.28%.
(205) 5. The brittleness and sweetness of can-like radish treated with the 5000 times diluted VdAL are improved, which indicates that the can-like radish treated with VdAL, has strong palatability.
(206) Experiments prove that the biological agent of VdAL according to the present invention, wherein VdAL is an active ingredient, can improve the drought resistance of plant:
(207) The plant treated with a solution having VdAL mass percentage concentration of 1-2% all die on the 15-20.sup.th day of water control, the plant not treated with VdAL all die on the 10-12.sup.th day of water control, and the dead time of the plant treated with the solution having VdAL mass percentage concentration of 1-2% is obviously longer than that of the plant not treated with VdAL. According to the re-watering treatment on the plant treated with a solution having VdAL mass percentage concentration of 0.2-2% after watering is stopped, the chlorophyll content of plant treated with VdAL is 34.5-34.4 SPAD, which is remarkably higher than that of plant not treated with VdAL; and the re-watering index of plant treated with VdAL is 0.43-0.52, which is remarkably higher than that of plant not treated with VdAL. It indicates that VdAL having a proper concentration can improve the drought resistance of wheat.
(208) Experiments prove that the biological agent of VdAL according to the present invention, wherein VdAL is an active ingredient, can promote coloration of plant fruit:
(209) 1. The color of cherry treated with the 5000 times diluted VdAL is significantly redder than that of untreated cherry, which indicated that VdAL can promote coloration of cherry.
(210) 2. After grape is treated with the 3000 times diluted VdAL, the color of grape is brighter.
(211) Experiments prove that protein VdAL and the biological agent thereof in the present invention can promote plant growth, improve the yield of plant, improve quality of plant product and palatability of plant fruit, improve the drought resistance of plant, promote coloration of plant fruit, and promote t fruit maturity.