CN107173393B - Microbial inoculum for preventing and treating diseases before and after strawberry harvest and promoting growth, preparation method and application - Google Patents
Microbial inoculum for preventing and treating diseases before and after strawberry harvest and promoting growth, preparation method and application Download PDFInfo
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Abstract
The invention discloses a microbial inoculum for preventing and treating postharvest diseases of strawberries before and after harvest and promoting growth, which comprises the following raw materials in percentage by mass: 85-95% of mixed zymocyte liquid, 0.05-0.1% of Tween 80, 0.2-0.5% of calcium chloride, 0.1-0.5% of sodium silicate, 0.75-2% of chitosan and the balance of water; the mixed zymophyte liquid is prepared from a meiqi yeast Y-3 zymophyte liquid, a burkholderia B-1 zymophyte liquid and an enterobacter B-6-1 zymophyte liquid according to the proportion of 1: 1-2: 0.5-1 by mass. In addition, the invention also provides a preparation method and application of the microbial inoculum for preventing and treating postharvest diseases of strawberries before harvest and promoting growth. The bactericide disclosed by the invention is sprayed before picking, so that the incidence rate of diseases after the strawberries are effectively reduced, the rotting rate of the strawberries after the strawberries are picked is reduced, the growth of the strawberries is promoted, and a new way is provided for green production and fresh keeping of the strawberries.
Description
Technical Field
The invention belongs to the technical field of microorganisms, and particularly relates to a microbial inoculum for preventing and treating postharvest diseases of strawberries before and after harvest and promoting growth, and a preparation method and application thereof.
Background
The strawberry belongs to the genus strawberry of the family Rosaceae, the fruit color is bright, the pulp is sour and sweet, and the nutrition and economic value are high. However, because the fruit peel is delicate, mechanical injury is easy to occur, and physiological disorder, water loss and pathogenic bacteria infection after picking are easy to cause. In the process of storing or storing and transporting the strawberries after picking, most of pathogenic bacteria causing fruit rot come from the field, so that effective measures are taken in the field to reduce the bacteria carrying amount on the surfaces of the picked fruits, and the method plays an important role in reducing the rot after picking.
At present, the main method for reducing rot of strawberries before and after picking is to use chemical bactericides which can threaten the environment and human health and bring the problems of drug resistance of pathogenic bacteria and the like, so that the application range of the strawberries is more and more limited, and other prevention and control methods are inevitably found.
Disclosure of Invention
The microbial inoculum for preventing and treating the diseases of the strawberries before and after picking and promoting the growth solves the problems that the strawberries are easy to generate the diseases before picking and are easy to rot after picking.
The invention aims to provide a microbial inoculum for preventing and treating postharvest diseases of strawberries before and after harvest and promoting growth, which comprises the following raw materials in percentage by mass: 85-95% of mixed zymocyte liquid, 0.05-0.1% of Tween 80, 0.2-0.5% of calcium chloride, 0.1-0.5% of sodium silicate, 0.75-2% of chitosan and the balance of water to 100%;
the total number of bacterial colonies in the mixed zymophyte liquid is 1.0 multiplied by 106～1.0×1011cfu/ml；
The mixed zymophyte liquid is prepared from Metschnikowia zizyphila Y-3 zymophyte liquid, Burkholderia B-1(Burkholderia contaminans B-1) zymophyte liquid and Enterobacter B-6-1(Enterobacter cowanii B-6-1) zymophyte liquid according to the weight ratio of 1: 1-2: 0.5-1 by mass;
wherein the Metschnikowia zizyphhicola Y-3 zymocyte liquid is obtained by culturing a Metschnikowia zizyphhicola Y-3 pure strain to the late logarithmic phase; the Burkholderia B-1 zymocyte liquid is obtained by culturing a pure Burkholderia B-1 strain to the late logarithmic phase; the enterobacter B-6-1 zymocyte liquid is obtained by culturing an enterobacter B-6-1 pure strain to the late logarithmic phase.
Preferably, the microbial inoculum for preventing and treating postharvest diseases of strawberries before and after harvest and promoting growth comprises the following raw materials in percentage by mass: 90% of mixed zymophyte liquid, 0.08% of Tween 80, 0.3% of calcium chloride, 0.3% of sodium silicate, 1% of chitosan and the balance of water to 100%;
the total number of bacterial colonies in the mixed zymophyte liquid is 1.0 multiplied by 108cfu/ml；
The mixed zymophyte liquid is prepared from Metschnikowia zizyphila Y-3 zymophyte liquid, Burkholderia B-1 zymophyte liquid and enterobacter B-6-1 zymophyte liquid according to the proportion of 2: 2: 1, and mixing the components in a mass ratio of 1.
The second purpose of the invention is to provide a preparation method of a microbial inoculum for preventing and treating postharvest diseases of strawberries before and after harvest and promoting growth, which comprises the following steps:
step 1, taking out a pure strain of Metschnikowia zizyphhicola Y-3 at-80 ℃, performing streak culture on a PDA plate for 48h at 28 ℃, then selecting a single colony in an NYDB liquid culture medium, and culturing for 72h at an oscillation speed of 200r/min to obtain an initial zymogen liquid of the Metschnikowia zizyphhicola Y-3;
taking out the pure bacterial strain of the Burkholderia B-1 at minus 80 ℃, performing streak culture on a PDA (personal digital Assistant) plate at 28 ℃ for 48h, picking a single bacterial colony in a YSP (YSP) liquid culture medium, and culturing at an oscillation speed of 200r/min for 24h to obtain an initial zymogen liquid of the Burkholderia B-1;
taking out a pure strain of the enterobacter B-6-1 at the temperature of minus 80 ℃, performing streak culture on a PDA (personal digital assistant) flat plate at the temperature of 28 ℃ for 48 hours, selecting a single colony in an LB (LB) liquid culture medium, and performing culture at the oscillation speed of 200r/min for 24 hours to obtain an initial zymogen liquid of the enterobacter B-6-1;
step 2, placing the initial zymogen liquid of Metschnikowia zizyphilia Y-3 in NYDB liquid culture medium, and then culturing at 28 ℃ at the oscillation speed of 200r/min to the end of logarithm to obtain the Metschnikowiazizyphilia Y-3 zymogen liquid;
placing the initial zymophyte liquid of the Burkholderia B-1 in an NYDB liquid culture medium, and then culturing at 28 ℃ at an oscillation speed of 200r/min to the end of logarithm to obtain the Burkholderia B-1 zymophyte liquid;
placing the initial zymocyte liquid of the enterobacter B-6-1 in an NYDB liquid culture medium, and then culturing at 28 ℃ at an oscillation speed of 200r/min to the end of logarithm to obtain the zymocyte liquid of the enterobacter B-6-1;
step 3, mixing the Metschnikowia zizyphila Y-3 zymocyte liquid, the Burkholderia B-1 zymocyte liquid and the enterobacter B-6-1 zymocyte liquid obtained in the step 2 according to the ratio of 1: 1-2: mixing the materials in a mass ratio of 0.5-1 to obtain a mixed zymophyte liquid;
step 4, weighing 90% of the mixed zymophyte liquid, 0.05% -0.1% of tween 80, 0.2-0.5% of calcium chloride, 0.1-0.5% of sodium silicate, 0.75% -2% of chitosan and the balance of water to make up 100%;
and 5, adding the Tween 80, the calcium chloride, the sodium silicate, the chitosan and the water which are weighed in the step 4 into the mixed fermentation bacteria liquid weighed in the step 4, and uniformly mixing to obtain the microbial inoculum for preventing and treating postharvest diseases of the strawberries and promoting growth.
Preferably, the following components are contained in each liter of the YSP liquid culture medium: 10g of peptone, 5g of yeast extract, 10g of glucose and the balance of water.
Preferably, the following components are contained in each liter of the NYDB liquid culture medium: 8g of beef extract, 5g of yeast extract, 10g of glucose and the balance of water.
Preferably, the LB liquid culture medium comprises the following components per liter: 10g of tryptone, 5g of yeast extract, 10g of NaCl and the balance of water.
The third purpose of the invention is to provide the application of the microbial inoculum for preventing and controlling the pre-harvest and post-harvest diseases of the strawberries and promoting the growth of the strawberries in the prevention and control of the pre-harvest and post-harvest diseases of the strawberries.
Microorganism deposit information description:
metschnikowia zizyphhicola Y-3: has been preserved in China center for type culture Collection, address: the preservation number of Wuhan university in Wuhan City of China is CCTCC NO. M2016564, and the preservation date is as follows: 2016, 10 months and 17 days.
Burkholderia B-1(Burkholderia contaminans B-1): has been preserved in China center for type culture Collection, address: the preservation number of Wuhan university in Wuhan City of China is CCTCC NO. M2014076, and the preservation date is as follows: 3, month and 7 days 2014.
Enterobacter sp.B-6-1 (Enterobacter cowanii B-6-1): has been preserved in China center for type culture Collection, address: the preservation number of Wuhan university in Wuhan City of China is CCTCC NO. M2012398, and the preservation date is as follows: 10/11/2012.
Wherein, the Burkholderia B-1 is disclosed in Chinese patent CN 104109645B entitled No. of granted publication No. 10/12/2016, bacterial strains and biocontrol microbial inoculum for preventing and treating postharvest diseases of fruits and vegetables;
enterobacter B-6-1 is disclosed in the Chinese patent ' biological antagonistic strain for preventing and treating postharvest diseases of fruits and vegetables ', its preparation method and application ' with the grant publication number CN 103952327B granted at 27.4.4.2016.
Compared with the prior art, the invention has the beneficial effects that:
1) the bactericide for preventing and treating postharvest diseases of strawberries and promoting growth, provided by the invention, is sprayed before the strawberries are picked, so that the incidence rate of postharvest diseases of strawberries can be effectively reduced, and the growth of strawberries can be promoted.
2) The bactericide for preventing and treating postharvest diseases of strawberries before and after picking and promoting growth, which is provided by the invention, is sprayed after the strawberries are picked, so that the bacterial carrying quantity on the surfaces of the strawberries can be effectively reduced, the rotting rate of the strawberries after the strawberries are picked can be reduced, and a new way is provided for green production and fresh keeping of the strawberries.
Drawings
FIG. 1 is a result diagram showing the influence of the pre-harvest and post-harvest disease prevention and growth promotion microbial inoculum prepared in the pre-harvest spraying example 2 on gray mold of strawberry fruits;
FIG. 2 is a result chart showing the influence of the microbial inoculum for preventing and treating postharvest diseases and promoting growth of strawberries on the natural decay of strawberry fruits, which is prepared in the pre-harvest spraying example 2.
Detailed Description
In order to make the technical solutions of the present invention better understood and enable those skilled in the art to practice the present invention, the following embodiments are further described, but the present invention is not limited to the following embodiments.
The experimental methods described in the examples of the present invention are all conventional methods unless otherwise specified.
Example 1
A microbial inoculum for preventing and treating postharvest diseases of strawberries before and after harvest and promoting growth comprises the following raw materials in percentage by mass: 85% of mixed zymocyte liquid, 0.05% of Tween 80, 0.5% of calcium chloride, 0.1% of sodium silicate, 2% of chitosan and the balance of water to 100%;
the total number of colonies in the mixed fermentation broth was 1.0X 106cfu/ml；
The mixed zymophyte liquid consists of Metschnikowia zizyphila Y-3 zymophyte liquid, Burkholderia B-1 zymophyte liquid and enterobacter B-6-1 zymophyte liquid according to the weight ratio of 1: 1.5: 0.8 by mass ratio;
wherein the Metschnikowia zizyphhicola Y-3 zymocyte liquid is obtained by culturing a Metschnikowia zizyphhicola Y-3 pure strain to the late logarithmic phase; the Burkholderia B-1 zymocyte liquid is obtained by culturing a pure Burkholderia B-1 strain to the late logarithmic phase; the Enterobacter B-6-1 zymocyte liquid is obtained by culturing the pure Enterobacter B-6-1 strain to the late logarithmic phase.
The specific implementation steps are as follows:
step 1, taking out a pure strain of Metschnikowia zizyphhicola Y-3 at-80 ℃, performing streak culture on a PDA plate for 48h at 28 ℃, then selecting a single colony in an NYDB liquid culture medium, and culturing for 72h at an oscillation speed of 200r/min to obtain an initial zymogen liquid of the Metschnikowia zizyphhicola Y-3;
taking out the pure bacterial strain of the Burkholderia B-1 at minus 80 ℃, performing streak culture on a PDA (personal digital Assistant) plate at 28 ℃ for 48h, picking a single bacterial colony in a YSP (YSP) liquid culture medium, and culturing at an oscillation speed of 200r/min for 24h to obtain an initial zymogen liquid of the Burkholderia B-1;
taking out a pure strain of the enterobacter B-6-1 at the temperature of minus 80 ℃, performing streak culture on a PDA (personal digital assistant) flat plate at the temperature of 28 ℃ for 48 hours, selecting a single colony in an LB (LB) liquid culture medium, and performing culture at the oscillation speed of 200r/min for 24 hours to obtain an initial zymogen liquid of the enterobacter B-6-1;
wherein, each liter of YSP liquid culture medium comprises the following components: 10g of peptone, 5g of yeast extract, 10g of glucose and the balance of water;
wherein, each liter of NYDB liquid culture medium contains the following components: 8g of beef extract, 5g of yeast extract, 10g of glucose and the balance of water;
wherein, each liter of LB liquid culture medium comprises the following components: 10g of tryptone, 5g of yeast extract, 10g of NaCl10g and the balance of water;
step 2, placing the initial zymogen liquid of Metschnikowia zizyphilia Y-3 in NYDB liquid culture medium, and then culturing at 28 ℃ at the oscillation speed of 200r/min to the end of logarithm to obtain the Metschnikowiazizyphilia Y-3 zymogen liquid;
placing the initial zymophyte liquid of the Burkholderia B-1 in an NYDB liquid culture medium, and then culturing at 28 ℃ at an oscillation speed of 200r/min to the end of logarithm to obtain the Burkholderia B-1 zymophyte liquid;
placing the initial zymocyte liquid of the enterobacter B-6-1 in an NYDB liquid culture medium, and then culturing at 28 ℃ at an oscillation speed of 200r/min to the end of logarithm to obtain the zymocyte liquid of the enterobacter B-6-1;
step 3, mixing the Metschnikowia zizyphila Y-3 zymocyte liquid, the Burkholderia B-1 zymocyte liquid and the enterobacter B-6-1 zymocyte liquid obtained in the step 2 according to the ratio of 1: 1.5: mixing according to the mass ratio of 0.8 to obtain mixed zymophyte liquid;
step 4, weighing 85% of the mixed zymophyte liquid, 0.05% of tween 80, 0.5% of calcium chloride, 0.1% of sodium silicate and 2% of chitosan by mass percent, and supplementing the balance of water to 100%;
and 5, adding the Tween 80, the calcium chloride, the sodium silicate, the chitosan and the water which are weighed in the step 4 into the mixed fermentation bacteria liquid weighed in the step 4, and uniformly mixing to obtain the microbial inoculum for preventing and treating postharvest diseases of the strawberries and promoting growth.
Example 2
A microbial inoculum for preventing and treating postharvest diseases of strawberries before and after harvest and promoting growth comprises the following raw materials in percentage by mass: 90% of mixed zymophyte liquid, 0.08% of Tween 80, 0.3% of calcium chloride, 0.3% of sodium silicate, 1% of chitosan and the balance of water to 100%;
the total number of colonies in the mixed fermentation broth was 1.0X 108cfu/ml；
The mixed zymophyte liquid is prepared from Metschnikowia zizyphila Y-3 zymophyte liquid, Burkholderia B-1 zymophyte liquid and Enterobacter B-6-1 zymophyte according to the proportion of 1: 1: 0.5 by mass ratio.
The specific implementation steps are as follows:
step 1, taking out a pure strain of Metschnikowia zizyphhicola Y-3 at the temperature of 80 ℃, performing streak culture on a PDA plate for 48h at the temperature of 28 ℃, then selecting a single colony in an NYDB liquid culture medium, and culturing for 72h at the oscillation speed of 200r/min to obtain an initial zymogen liquid of the Metschnikowia zizyphhicola Y-3;
taking out the pure bacterial strain of the Burkholderia B-1 at minus 80 ℃, performing streak culture on a PDA (personal digital Assistant) plate at 28 ℃ for 48h, picking a single bacterial colony in a YSP (YSP) liquid culture medium, and culturing at an oscillation speed of 200r/min for 24h to obtain an initial zymogen liquid of the Burkholderia B-1;
taking out a pure strain of the enterobacter B-6-1 at the temperature of minus 80 ℃, performing streak culture on a PDA (personal digital assistant) flat plate at the temperature of 28 ℃ for 48 hours, selecting a single colony in an LB (LB) liquid culture medium, and performing culture at the oscillation speed of 200r/min for 24 hours to obtain an initial zymogen liquid of the enterobacter B-6-1;
wherein the NYDB liquid culture medium, YSP liquid culture medium and LB liquid culture medium are the same as in example 1;
step 2, placing the initial zymogen liquid of Metschnikowia zizyphilia Y-3 in NYDB liquid culture medium, and then culturing at 28 ℃ at the oscillation speed of 200r/min to the end of logarithm to obtain the Metschnikowiazizyphilia Y-3 zymogen liquid;
placing the initial zymophyte liquid of the Burkholderia B-1 in an NYDB liquid culture medium, and then culturing at 28 ℃ at an oscillation speed of 200r/min to the end of logarithm to obtain the Burkholderia B-1 zymophyte liquid;
placing the initial zymocyte liquid of the enterobacter B-6-1 in an NYDB liquid culture medium, and then culturing at 28 ℃ at an oscillation speed of 200r/min to the end of logarithm to obtain the zymocyte liquid of the enterobacter B-6-1;
step 3, mixing the Metschnikowia zizyphila Y-3 zymocyte liquid, the Burkholderia B-1 zymocyte liquid and the enterobacter B-6-1 zymocyte liquid obtained in the step 2 according to the ratio of 1: 1: mixing according to the mass ratio of 0.5 to obtain mixed zymophyte liquid;
step 4, weighing 90% of the mixed zymophyte liquid, 0.08% of tween 80, 0.3% of calcium chloride, 0.3% of sodium silicate and 1% of chitosan by mass percent, and supplementing the balance of water to 100%;
and 5, adding the Tween 80, the calcium chloride, the sodium silicate, the chitosan and the water which are weighed in the step 4 into the mixed fermentation bacteria liquid weighed in the step 4, and uniformly mixing to obtain the microbial inoculum for preventing and treating postharvest diseases of the strawberries and promoting growth.
Example 3
A microbial inoculum for preventing and treating postharvest diseases of strawberries before and after harvest and promoting growth comprises the following raw materials in percentage by mass: 95% of mixed zymophyte liquid, 0.1% of Tween 80, 0.2% of calcium chloride, 0.5% of sodium silicate, 0.75% of chitosan and the balance of water for 100%;
the total number of bacterial colonies in the mixed zymophyte liquid is 1.0 multiplied by 1011cfu/ml；
The mixed zymophyte liquid is prepared from Metschnikowia zizyphila Y-3 zymophyte liquid, Burkholderia B-1 zymophyte liquid and enterobacter B-6-1 zymophyte liquid according to the proportion of 1: 2: 1 by mass ratio;
the specific implementation steps are as follows:
step 1, taking out a pure strain of Metschnikowia zizyphhicola Y-3 at-80 ℃, performing streak culture on a PDA plate for 48h at 28 ℃, then selecting a single colony in an NYDB liquid culture medium, and culturing for 72h at an oscillation speed of 200r/min to obtain an initial zymogen liquid of the Metschnikowia zizyphhicola Y-3;
taking out the pure bacterial strain of the Burkholderia B-1 at minus 80 ℃, performing streak culture on a PDA (personal digital Assistant) plate at 28 ℃ for 48h, picking a single bacterial colony in a YSP (YSP) liquid culture medium, and culturing at an oscillation speed of 200r/min for 24h to obtain an initial zymogen liquid of the Burkholderia B-1;
taking out a pure strain of the enterobacter B-6-1 at the temperature of minus 80 ℃, performing streak culture on a PDA (personal digital assistant) flat plate at the temperature of 28 ℃ for 48 hours, selecting a single colony in an LB (LB) liquid culture medium, and performing culture at the oscillation speed of 200r/min for 24 hours to obtain an initial zymogen liquid of the enterobacter B-6-1;
wherein the NYDB liquid culture medium, YSP liquid culture medium and LB liquid culture medium are the same as in example 1;
step 2, placing the initial zymogen liquid of Metschnikowia zizyphilia Y-3 in NYDB liquid culture medium, and then culturing at 28 ℃ at the oscillation speed of 200r/min to the end of logarithm to obtain the Metschnikowiazizyphilia Y-3 zymogen liquid;
placing the initial zymophyte liquid of the Burkholderia B-1 in an NYDB liquid culture medium, and then culturing at 28 ℃ at an oscillation speed of 200r/min to the end of logarithm to obtain the Burkholderia B-1 zymophyte liquid;
placing the initial zymocyte liquid of the enterobacter B-6-1 in an NYDB liquid culture medium, and then culturing at 28 ℃ at an oscillation speed of 200r/min to the end of logarithm to obtain the zymocyte liquid of the enterobacter B-6-1;
step 3, mixing the Metschnikowia zizyphila Y-3 zymocyte liquid, the Burkholderia B-1 zymocyte liquid and the enterobacter B-6-1 zymocyte liquid obtained in the step 2 according to the ratio of 1: 2: 1 to obtain mixed zymocyte liquid;
step 4, weighing 95% of mixed zymocyte liquid, 0.1% of Tween 80, 0.2% of calcium chloride, 0.5% of sodium silicate, 0.75% of chitosan and the balance of water to make up 100%;
and 5, adding the Tween 80, the calcium chloride, the sodium silicate, the chitosan and the water which are weighed in the step 4 into the mixed fermentation bacteria liquid weighed in the step 4, and uniformly mixing to obtain the microbial inoculum for preventing and treating postharvest diseases of the strawberries and promoting growth.
The effect of the present invention will be further explained by the application of the microbial inoculum for preventing and treating the postharvest diseases of strawberries and promoting growth.
Since the performance parameters of the strawberries measured after the microbial inoculum prepared in the embodiments 1 to 3 is used for spraying strawberries are similar, only the performance parameters measured after the strawberry is sprayed with the microbial inoculum preferred in the embodiment 2 are taken as an illustration.
Experiment for preventing and treating strawberry diseases by using the microbial inoculum for preventing and treating diseases before and after strawberry harvest and promoting growth in example 2
Experimental materials: strawberry (variety is "Hongyan") is planted in the east yang town of Ulmus City of Shanxi province, and no chemical bactericide is used.
And (3) comparison: sterile water spray was used.
Treating a sample: the microbial inoculum prepared in example 2 for preventing and controlling postharvest diseases of strawberries and promoting growth is used.
The implementation method comprises the following steps: respectively using a manual sprayer to spray strawberries 6d, 3d and 0d before picking until liquid drops drip, repeatedly spraying for 3 times in each spraying period, repeatedly spraying 40 strawberries in each spraying period, conveying the sprayed strawberry fruits back to a laboratory after the strawberries are picked in the mature period, selecting the fruits with consistent color and luster and no mechanical injury for experiment, wherein the experimental results are as follows:
1. control of gray mold of strawberry fruits by spraying microbial inoculum before picking
Collecting the above strawberry fruits at a concentration of 1 × 105Dipping the spores/ml of botrytis cinerea suspension for 10s, drying the suspension in sterile air, putting the suspension into a sterilized plastic box, adding a plastic bag for moisturizing, sequentially placing the box at 6 ℃ for 15 days and 16 ℃ for 7 days, recording the disease incidence, and repeating the treatment of each batch for 3 times, wherein the specific experimental result is shown in figure 1.
As can be seen from FIG. 1, the incidence of Botrytis cinerea of strawberry fruits treated with the microbial inoculum after being stored for 7 days at 16 ℃ is 32.23%, while the incidence of Botrytis cinerea of strawberry fruits of the control is 93.87%; the disease condition of the strawberry fruit botrytis cinerea stored at 6 ℃ for 15 days is similar to that of the strawberry fruit botrytis cinerea stored at 16 ℃ for 7 days, and the disease rate of the strawberry fruit botrytis cinerea treated by spraying the microbial inoculum for 3 times is obviously reduced compared with a control sample, so that the disease rate of the strawberry fruit botrytis cinerea after being collected can be well controlled by spraying the microbial inoculum compared with the strawberry control sample sprayed with sterile water.
2. Influence of spraying microbial inoculum before picking on natural rot of strawberry fruits
The harvested strawberry fruits are put into a plastic box, the plastic box is placed at 6 ℃ for 15 days and 16 ℃ for 7 days in a moisture-preserving mode, the rotting rate of the fruits is recorded, each batch of processing is repeated for 3 times, and specific experimental results are shown in figure 2.
As can be seen from figure 2, after the microbial inoculum is sprayed before picking, the rotting rate of the strawberry fruits treated by the microbial inoculum is obviously reduced at two different temperatures, and the rotting rate is 24.14% at 6 ℃ and 15 days and 20.72% at 16 ℃ and 7 days after treatment. The decay rate of the control was 78.98% at 16 ℃ and 7d and 76.73% at 6 ℃ and 15 d. Therefore, the strawberries can be well prevented from rotting and the rotting rate of the strawberries can be reduced by spraying the compound microbial inoculum.
3. Influence of spraying microbial inoculum before picking on postharvest quality of strawberry fruits
The color difference values of the fruit surfaces were measured by a CR-400 type color difference meter (Minolta, japan), where L denotes brightness, a negative value denotes green light, a positive value denotes red light, H denotes hue angle, the color difference values were measured at the equator of the fruit, and the influence of the spray of the microbial inoculum before harvest on the color difference of the strawberry fruit surfaces after harvest is shown in table 1.
TABLE 1 Effect of pre-harvest treatment with Complex microbial inoculum on color change of strawberries after harvest
As can be seen from Table 1, the brightness values of the samples treated by the complex microbial inoculum under two different temperature conditions are reduced after the storage period. Although the L values of the treated samples were slightly higher than the control samples after storage at 7d and 15d at both temperatures, the difference was not significant compared to the control samples. and the a value and the H value are also similar to each other, the a value of each treatment sample is generally increased after storage, the H value is generally reduced, the color of the fruit is further turned to red, and the difference between the treatment sample and the control sample is not obvious, so that the influence of the spraying of the microbial inoculum before the sampling on the color of the strawberry fruit in the storage period is not obvious.
The effect of spraying the microbial inoculum on the strawberry quality before harvest is shown in table 2.
TABLE 2 Effect of pre-harvest treatment with Complex microbial inoculum on post-harvest quality of strawberries
The influence of the spraying of the microbial inoculum on the quality of the strawberries before picking is shown in table 2, and the weight loss rate of the strawberries after being stored for 7 days at 16 ℃ is larger than that of the strawberries after being stored for 15 days at 6 ℃, but the strawberry fruit weight loss rate has no obvious difference under the same temperature condition. In addition, the spraying of the microbial inoculum before picking can delay the reduction of fruit hardness, SSC, titratable acid and Vc content to a certain extent.
Experiment for controlling postharvest diseases of strawberries and promoting growth of strawberries by using microbial inoculum for controlling postharvest diseases of strawberries in example 2
Experimental materials: strawberry (variety is "Hongyan") is planted in the east yang town of Ulmus City of Shanxi province, and no chemical bactericide is used.
And (3) comparison: sterile water spray was used.
Treating a sample: the microbial inoculum prepared in example 2 for preventing and controlling postharvest diseases of strawberries and promoting growth is used.
The experimental process comprises the following steps: and (3) spraying the strawberry plants in the plots with serious powdery mildew in the full bearing period of the strawberries (ten days of 4 months), and spraying the fungicide 1 time every 2 days for 3 times. When the strawberry spray is carried out, a manual sprayer is used for spraying the strawberries until liquid drops are formed, 120 strawberries are repeatedly sprayed each time, and the control is not treated. And after 10d of spraying the microbial inoculum for the last time, observing and recording the growth conditions of the control and treated strawberry plants, and recording the plant height, the fresh weight of the overground part, the number of leaves of each plant and the occurrence condition of powdery mildew of the strawberry plants.
1. Growth promoting effect of microbial inoculum on strawberries
The growth promoting effect on strawberries after the compound microbial inoculum is sprayed is shown in table 3.
TABLE 3 Effect of inoculant treatment on strawberry seedling growth
| Treatment of | Plant height (cm) | Fresh weight of plant | Number of leaves per plant |
| Processing sample | 24.26±0.27 | 103.59±2.42 | 7.53±0.28 |
| Control sample | 19.56±0.43 | 89.42±2.83 | 5.31±0.42 |
As can be seen from Table 3, spraying 3 times of microbial inoculum before harvest has a certain promotion effect on the growth of strawberry plants. Compared with a control plant, the strawberry plant sprayed for 3 times is stronger, the leaves are dark green, the growth vigor is obviously superior to that of the control plant, and the plant height, the fresh weight and the number of the leaves of the single plant sprayed for 3 times before harvest are obviously increased.
2. Inhibiting effect of microbial inoculum on powdery mildew
After spraying the bactericide agent for 10 days, the strawberries after the treatment and the control enter the red turning period are all harvested, the morbidity of the powdery mildew of the strawberries and the powdery mildew of plants are respectively counted, and the experimental results are shown in table 4.
TABLE 4 Effect of microbial inoculum treatment on strawberry seedling powdery mildew
| Treatment of | Incidence of disease in plants | Fruit morbidity |
| Processing sample | 12.97±1.42 | 56.43±2.18 |
| Control sample | 27.52±1.35 | 29.08±1.53 |
As can be seen from Table 4, the plant morbidity and fruit morbidity of the treated sample are obviously lower than those of the control sample, and the strawberry seedlings have obvious effect of inhibiting the powdery mildew morbidity after the compound microbial inoculum is sprayed.
While preferred embodiments of the present invention have been described, additional variations and modifications in those embodiments may occur to those skilled in the art once they learn of the basic inventive concepts. Therefore, it is intended that the appended claims be interpreted as including preferred embodiments and all such alterations and modifications as fall within the scope of the invention.
It will be apparent to those skilled in the art that various changes and modifications may be made in the present invention without departing from the spirit and scope of the invention. Thus, it is intended that such changes and modifications be included within the scope of the appended claims and their equivalents.
Claims (6)
1. A preparation method of a microbial inoculum for preventing and treating postharvest diseases of strawberries before and after harvest and promoting growth comprises the following raw materials in percentage by mass: 85-95% of mixed zymophyte liquid, 0.05-0.1% of Tween 80, 0.2-0.5% of calcium chloride, 0.1-0.5% of sodium silicate, 0.75-2% of chitosan and the balance of water to 100%;
the total number of bacterial colonies in the mixed zymophyte liquid is 1.0 multiplied by 106～1.0×1011cfu/mL；
The mixed zymocyte liquid consists of a Metschnikowia zizyphicola Y-3 zymocyte liquid, a Burkholderia B-1(Burkholderia continans B-1) zymocyte liquid and an Enterobacter B-6-1(Enterobacter cowanii B-6-1) zymocyte liquid, and the mass of the Metschnikowia zizyphicola Y-3 zymocyte liquid is as follows: the quality of the Burkholderia B-1 zymocyte liquid is as follows: the mass of the enterobacter B-6-1 zymocyte liquid is 1: 1-2: 0.5 to 1;
wherein the Meiqi yeast Y-3 zymocyte liquid is obtained by culturing the Meiqi yeast Y-3 pure strain to the late logarithmic phase; the Burkholderia B-1 zymocyte liquid is obtained by culturing a pure Burkholderia B-1 strain to the late logarithmic phase; the enterobacter B-6-1 zymocyte liquid is obtained by culturing an enterobacter B-6-1 pure strain to the late logarithmic phase;
the method is characterized by comprising the following steps:
step 1, taking out a pure strain of the Metschnikowia Y-3 at-80 ℃, performing streak culture on a PDA (personal digital Assistant) plate for 48 hours at 28 ℃, and then selecting a single colony to be cultured in an NYDB liquid culture medium for 72 hours at an oscillation speed of 200r/min to obtain an initial zymogen liquid of the Metschnikowia Y-3;
taking out the pure bacterial strain of the Burkholderia B-1 at minus 80 ℃, performing streak culture on a PDA (personal digital Assistant) plate at 28 ℃ for 48h, selecting a single bacterial colony, and culturing the single bacterial colony in a YSP (YSP) liquid culture medium at an oscillation speed of 200r/min for 24h to obtain an initial zymogen liquid of the Burkholderia B-1;
taking out a pure strain of the enterobacter B-6-1 at the temperature of minus 80 ℃, performing streak culture on a PDA (PDA) plate at the temperature of 28 ℃ for 48 hours, and then selecting a single colony to be cultured in an LB (LB) liquid culture medium at the oscillation speed of 200r/min for 24 hours to obtain an initial zymogen liquid of the enterobacter B-6-1;
step 2, placing the initial zymocyte liquid of the Metschnikowia Y-3 in an NYDB liquid culture medium, and then culturing at the temperature of 28 ℃ for 72 hours at the oscillation speed of 200r/min to obtain the zymocyte liquid of the Metschnikowia Y-3;
placing the initial zymophyte liquid of the Burkholderia B-1 in an NYDB liquid culture medium, and then culturing at 28 ℃ at an oscillation speed of 200r/min for 24h to obtain the Burkholderia B-1 zymophyte liquid;
putting the initial zymophyte liquid of the enterobacter B-6-1 into an NYDB liquid culture medium, and then culturing at the temperature of 28 ℃ and the oscillation speed of 200r/min for 24h to obtain the zymophyte liquid of the enterobacter B-6-1;
step 3, mixing the Metschnikowia Y-3 fermented bacterial liquid, the Burkholderia B-1 fermented bacterial liquid and the Enterobacter B-6-1 fermented bacterial liquid obtained in the step 2 according to the mass of the Metschnikowia Y-3 fermented bacterial liquid: the quality of the Burkholderia B-1 zymocyte liquid is as follows: the mass of the enterobacter B-6-1 zymocyte liquid is 1: 1-2: mixing the materials in a ratio of 0.5-1 to obtain a mixed zymophyte liquid;
step 4, weighing 85-95% of the mixed zymophyte liquid, 0.05-0.1% of Tween 80, 0.2-0.5% of calcium chloride, 0.1-0.5% of sodium silicate, 0.75-2% of chitosan and the balance of water to complement 100% for later use;
and 5, adding the Tween 80, the calcium chloride, the sodium silicate, the chitosan and the water which are weighed in the step 4 into the mixed fermentation bacteria liquid weighed in the step 4, and uniformly mixing to obtain the microbial inoculum for preventing and treating postharvest diseases of the strawberries and promoting growth.
2. The preparation method of the microbial inoculum for preventing and treating pre-harvest and post-harvest diseases of strawberries and promoting growth according to claim 1, wherein the microbial inoculum comprises the following raw materials in percentage by mass: 90% of mixed zymophyte liquid, 0.08% of Tween 80, 0.3% of calcium chloride, 0.3% of sodium silicate, 1% of chitosan and the balance of water to 100%;
the total number of bacterial colonies in the mixed zymophyte liquid is 1.0 multiplied by 108cfu/ml；
The mixed zymocyte liquid consists of a Metschnikowia Y-3 zymocyte liquid, a Burkholderia B-1 zymocyte liquid and an enterobacter B-6-1 zymocyte liquid, and the quality of the Metschnikowia Y-3 zymocyte liquid is as follows: the quality of the Burkholderia B-1 zymocyte liquid is as follows: the mass of the enterobacter B-6-1 zymocyte liquid is 1: 1: 0.5;
wherein the Meiqi yeast Y-3 zymocyte liquid is obtained by culturing the Meiqi yeast Y-3 pure strain to the late logarithmic phase; the Burkholderia B-1 zymocyte liquid is obtained by culturing a pure Burkholderia B-1 strain to the late logarithmic phase; the enterobacter B-6-1 zymocyte liquid is obtained by culturing an enterobacter B-6-1 pure strain to the late logarithmic phase.
3. The preparation method of the microbial inoculum for preventing and treating pre-harvest and post-harvest diseases of strawberries and promoting growth according to claim 1, wherein each liter of YSP liquid culture medium comprises the following components: 10g of peptone, 5g of yeast extract, 10g of glucose and the balance of water.
4. The preparation method of the microbial inoculum for preventing and treating pre-harvest and post-harvest diseases of strawberries and promoting growth according to claim 3, wherein each liter of the NYDB liquid culture medium contains the following components: 8g of beef extract, 5g of yeast extract, 10g of glucose and the balance of water.
5. The preparation method of the microbial inoculum for preventing and treating pre-harvest and post-harvest diseases of strawberries and promoting growth according to claim 4, wherein each liter of the LB liquid culture medium comprises the following components: 10g of tryptone, 5g of yeast extract, 10g of NaCl and the balance of water.
6. The application of the microbial inoculum prepared by the method of claim 1 for preventing and treating pre-harvest and post-harvest diseases of strawberries and promoting growth in the growth of strawberries and the prevention and treatment of pre-harvest and post-harvest diseases of strawberries.
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