CN107198782B - Method for killing ulcer disease pathogenic bacteria PSA after kiwi pollen is picked - Google Patents
Method for killing ulcer disease pathogenic bacteria PSA after kiwi pollen is picked Download PDFInfo
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- CN107198782B CN107198782B CN201710434050.4A CN201710434050A CN107198782B CN 107198782 B CN107198782 B CN 107198782B CN 201710434050 A CN201710434050 A CN 201710434050A CN 107198782 B CN107198782 B CN 107198782B
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2/00—Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor
- A61L2/02—Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor using physical phenomena
- A61L2/025—Ultrasonics
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2/00—Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor
- A61L2/02—Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor using physical phenomena
- A61L2/08—Radiation
- A61L2/081—Gamma radiation
Abstract
The invention discloses a post-harvest killer for kiwi fruit pollenA method of eradicating ulcer disease pathogenic PSA comprising the steps of: 1) subpackaging and sealing the picked kiwi fruit pollen in plastic bottles, placing the plastic bottles in an ultrasonic cleaner, carrying out ice-water bath, wherein the ultrasonic radiation power is 30-60W, and the radiation treatment time is 3-15 minutes; 2) immediately feeding the pollen after ultrasonic treatment into the container60Co-gamma ray irradiation is carried out in the irradiation bin, the irradiation dose is 400-1000 Gry, and the irradiation temperature is as follows: 25 +/-5 ℃; 3) storing the pollen after irradiation at-20 deg.C; 4) taking out pollen before pollination, and standing at room temperature for 3 hours; 5) and (3) fully mixing the pollen and the lycopodium clavatum powder according to the mass ratio of 1: 1-1: 0.4, and then pollinating the female flowers. Under reasonable process conditions, compared with untreated pollen, the activity of the pollen of an irradiation treatment sample is reduced by less than or equal to 40%, the plant fruit setting rate is greater than or equal to 92%, and the method is low in cost, good in effect and high in operability, and the application of the technology in the field of sterilization and survival after plant pollen collection is not seen.
Description
Technical Field
The invention belongs to a sterilization method of agricultural products, and particularly relates to a method for killing ulcer disease pathogenic bacteria PSA after kiwi pollen is picked, which is particularly suitable for post-picking sterilization infected with PSA kiwi pollen.
Background
Due to the rapid development of kiwi fruit industries in various countries in the world in recent years, along with the rapid increase of scale and increasingly remarkable contradiction between shortage of rural and agricultural labor force, the trend of using kiwi fruit commodity pollen is inevitable.
Most of kiwi pollen is produced from artificially-planted male plants, the spreading and latency of the kiwi canker are extremely strong, and the male plants are difficult to avoid infecting Pseudomonas Syringae (PSA), a pathogenic bacterium of the kiwi canker. Although some wild flowers are collected, the cross contamination problem also exists in the processing process. Thus, the rapid, large-area, trans-regional spread of kiwifruit canker by commercial pollen is highly likely[1]。
The kiwifruit canker is a low-temperature and high-humidity disease, and is a destructive bacterial disease seriously threatening the production of kiwifruit[2]The disease was first reported in New Zealand in 1973 and was discovered in California, USA and Jinggang county, Japan in 1980, and then rapidly spread to all large kiwifruit producing areas around the world. The outbreak of the kiwifruit canker disease of artificially cultivated kiwifruit is first reported in China in the eastern mountain peak forest farm of Shimen county of Hunan province in 1988[3]At present, with the continuous expansion of the artificial cultivation scale of Chinese kiwi fruit, the disease condition of kiwifruit canker is getting more and more serious, and the kiwifruit canker is listed as a national forest plant quarantine object[4]. And the commercial pollen has the hidden trouble of spreading PSA in a large area across regions. Therefore, a method for effectively killing canker pathogenic bacteria (PSA) carried by kiwi pollen and keeping the activity of the PSA to ensure production is developed. Has great significance for ensuring the sustainable development of the kiwi fruit industry.
The traditional physical sterilization and chemical sterilization methods are not suitable for pollen sterilization because the traditional physical sterilization and chemical sterilization methods are difficult to maintain pollen viability while killing pathogenic bacteria.
The irradiation technology (microwave, ultrasonic wave, ray) has been widely used for killing various microorganisms in medical health care products, animal feed, food, medicines and agricultural products, improving the sanitary quality and prolonging the shelf life[5]. In the sterilization irradiation technology, microwave irradiation generates a large amount of heat and is only suitable for articles insensitive to temperature; the penetration force of ultraviolet radiation is weak, and the ultraviolet radiation is suitable for surface sterilization; the gamma-ray irradiation has strong ray penetrating power, can effectively kill putrefying microorganisms and pathogenic bacteria, can sterilize while not damaging the external package, avoids secondary pollution, and does not generate heat effect and change heat effect when the irradiation sterilization is carried out at normal temperatureTemperature of the processed material (also known as cold working), sterilization method which is optimal especially for heat-sensitive articles[5][6]. A great deal of research shows that under the appropriate irradiation dose, the protein, lipid and carbohydrate in crops are only slightly changed during the irradiation process[7,8]。
The invention is sterilized by contrast heating, ultraviolet irradiation, microwave, single ultrasonic irradiation and single60Pollen germination method reported in Co-gamma ray irradiation sterilization reference literature[10]And PCR method[1]Detecting pollen, and finding that: firstly, the sample is irradiated by ultrasonic wave and then passes through60Co-gamma ray irradiation can be effectively reduced60The radiation dose of Co-gamma ray can kill PSA completely and keep pollen vitality.
Reference to the literature
[1]Vanneste J L,Giovanardi D,Yu J,et al.Detection of Pseudomonassyringae pv.actinidiae in kiwifruit pollen samples.[J].New Zealand PlantProtection,2011,64:246-251.
[2] The adaptability of the bacterial canker of Kiwi fruit in China is analyzed [ J ]. plant protection, 2016,42(2): 146-.
[3] The generation and prevention of ulcer of kiwi fruit is communicated with agricultural science and technology, 1988(12):175-17.
[4] Liu Qing gang, Chinese gooseberry canker occurrence rule and comprehensive prevention and treatment technology [ J ] Sichuan agricultural science and technology 2011(7) 38-39.
[5]Lambert P A.Radiation Sterilization[M]//Russell,Hugo&Ayliffe's:Principles and Practice of Disinfection,Preservation and Sterilization,5thEdition.Wiley‐Blackwell,2012:586-592.
[6] Harmine, Shihuilong, Wangfeng, et al. current research situation and application characteristics of electron beam food irradiation [ J ]. Nuclear agriculture Association, 2007,21(1):61-64.
[7] Zhujiating, von Min, Liuchunquan, and the like, radiation sterilization research on soybean protein powder [ J ] Nuclear agriculture bulletin, 2008,22(5): 645-.
[8] Influence of yellow wavelet, Mameihu, Liwenge on characteristics of egg protein liquid by radiation sterilization [ J ] agricultural engineering report, 2009,25(5): 244-.
[9] Studies on the sterilization effect of the tomato paste by using the new dun yuan, the yellow minium and the ultrasonic treatment in cooperation with the 60Co gamma irradiation [ J ] Guangdong agricultural science, 2011,38(4): 105-.
[10] Jiawenqing, Zhang Shaowei, Liu Lu Ying, etc. the influence of different culture media and storage methods on the pollen germination of the short peony [ J ]. southwest agro-journal, 2013,26(1): 338-.
Disclosure of Invention
The invention aims to provide a method for killing pathogenic bacteria of canker sore PSA after kiwi pollen is picked60Co-gamma ray irradiation can be effectively reduced60The radiation dose of Co-gamma ray can kill PSA completely and keep pollen vitality. The method has the advantages of low cost, good effect and high operability, and the application of the technology in the field of sterilization and keep-alive of the picked plant pollen is not seen.
The main technical problems of the invention are realized by adopting the following technical scheme: a method for killing canker pathogenic bacteria PSA after kiwi pollen is picked comprises the following steps:
1) subpackaging and sealing the picked kiwi fruit pollen in plastic bottles, placing the plastic bottles in an ultrasonic cleaner, carrying out ice-water bath, wherein the ultrasonic radiation power is 30-60W, and the radiation treatment time is 3-15 minutes;
2) immediately feeding the pollen after ultrasonic treatment into the container60Co-gamma ray irradiation is carried out in the irradiation bin, the irradiation dose is 400-1000 Gry, and the temperature is as follows: 25 +/-5 ℃;
3) storing the pollen after irradiation at-20 deg.C;
4) taking out pollen before pollination, and standing at room temperature for 3 hours;
5) and (3) fully mixing the pollen and the lycopodium clavatum powder according to the mass ratio of 1: 1-1: 0.4, and then pollinating the female flowers.
The ultrasonic radiation environment in the step 1) is as follows: sealing pollen in a plastic bottle, immersing in an ice-water mixture at 0-3 ℃, and carrying out ultrasonic radiation treatment.
The height of the plastic bottle in the step 1) is 3cm, and the thickness of the plastic bottle is 3 mm.
And finally, determining whether the sterilized kiwi fruit pollen is positive to PSA (prostate specific antigen) by adopting a PCR (polymerase chain reaction), and determining the activity of the sterilized pollen by adopting a pollen germination determination method.
Compared with the prior art, the invention has obvious advantages and excellent effects. According to the technical scheme, the kiwi fruit pollen is harvested, enriched and frozen at low temperature, taken out before pollination and controlled at a certain temperature, the pollen is subpackaged in plastic bottles, ultrasonic irradiation is firstly used, and then a certain dosage of pollen is carried out60Co-gamma ray is irradiated to achieve the aim of killing PSA carried by the kiwi fruit pollen. And (3) after irradiation sterilization, fully mixing the pollen with the lycopodium clavatum powder according to the mass ratio, and then pollinating the female flowers. Under reasonable process conditions, the pollen viability of the irradiation treated sample is reduced compared to untreated pollen.
Detailed Description
The following detailed description of the embodiments, features and effects of the method for killing pathogenic bacteria of canker sore PSA after picking kiwi pollen according to the present invention will be made in conjunction with the experiments and the preferred embodiments.
The infection of PSA is difficult to avoid in the large-scale production process of kiwi fruit commercial pollen. In order to avoid the hidden trouble that the commercial pollen spreads PSA when being used in a large area and across areas in the using process, the invention firstly compares the heat sterilization, the ultrasonic wave, the microwave and the60The kiwi pollen is treated by four modes of Co-gamma ray irradiation (detailed in Table 1), and the result shows that the sterilization is not thorough in the presence of ultrasonic waves, and the presumed reason is that the penetrating capacity is weak; although microwave can sterilize for a short time, the microwave has strong capability of killing pollen activity, and is presumed to be a heat effect reason;60although the Co-gamma ray irradiation effect is very obvious, the defect that the pollen activity is greatly reduced due to large dose irradiation exists, and the presumed reason is that the penetration capability is strong, the energy is large, and the control is difficult. Through a large number of control experiments, it was found that: firstly, the sample is irradiated by ultrasonic wave and then passes through60Co-gamma ray irradiation can effectively reduce the sterilization effect on the premise of achieving the sterilization effect60Co-gamma ray irradiation dose.
TABLE 1 Effect of different sterilization techniques on Kiwi fruit pollen PSA kill and viable pollen viability (n ═ 3)
Note: experimental scale: 50 g/time of pollen, pollen viability before sterilization: 47.9 percent.
The invention compares heat sterilization, single ultrasonic wave, microwave and60after Co-gamma ray irradiation sterilization effect, the sterilization capability and pollen activity are taken as sterilization process feasibility indexes, and ultrasonic treatment combination is screened out60The Co-gamma ray irradiation sterilization technology has very obvious effect, can achieve the effect of effectively killing PSA and can also inhibit the reduction of pollen activity to the maximum extent. The obtained process has strong operability, PSA infected by kiwi fruit pollen can be thoroughly killed, and the pollen activity is reduced by less than or equal to 40%.
A method for killing canker pathogenic bacteria PSA after kiwi pollen is picked comprises the following steps:
1) subpackaging and sealing the picked kiwi fruit pollen in plastic bottles, placing the plastic bottles in an ultrasonic cleaner, carrying out ice-water bath, wherein the ultrasonic radiation power is 30-60W, and the radiation treatment time is 3-15 minutes;
2) immediately feeding the pollen after ultrasonic treatment into the container60Co-gamma ray irradiation is carried out in the irradiation bin, the irradiation dose is 400-1000 Gry, and the temperature is as follows: 25 +/-5 ℃;
3) storing the pollen after irradiation at-20 deg.C;
4) taking out pollen before pollination, and standing at room temperature for 3 hours;
5) and (3) fully mixing the pollen and the lycopodium clavatum powder according to the mass ratio of 1: 1-1: 0.4, and then pollinating the female flowers.
The ultrasonic radiation environment in the step 1) is as follows: sealing pollen in a plastic bottle, immersing in an ice-water mixture at 0-3 ℃, and carrying out ultrasonic radiation treatment.
The height of the plastic bottle in the step 1) is 3cm, and the thickness of the plastic bottle is 3 mm.
And finally, determining whether the sterilized kiwi fruit pollen is positive to PSA (prostate specific antigen) by adopting a PCR (polymerase chain reaction), and determining the activity of the sterilized pollen by adopting a pollen germination determination method.
Example 1 (pollen sample one):
1) subpackaging the picked kiwi fruit pollen, sealing in a plastic bottle with the height of 3cm and the thickness of 3mm, 10 g/bottle, totaling 200 bottles, placing in an ultrasonic cleaner, performing ice-water bath with the power of 30W, and treating for 15 minutes;
2) immediately feeding the pollen after ultrasonic treatment into the container60Co-gamma ray irradiation is carried out in an irradiation bin, the irradiation dose is 1000Gry, and the temperature is 25 ℃;
3) storing the pollen after irradiation at-20 deg.C;
4) taking out pollen before pollination, and standing at room temperature for 3 hours;
5) fully mixing pollen and lycopodium clavatum powder according to the mass ratio of 1:0.4, and pollinating 13,881 female flowers on 50 kiwi trees;
6) the pollen viability is determined by a pollen germination method, the control pollen sample viability is 43.7%, the irradiation pollen sample viability is 19.8%, and the PSA detection is negative.
7) Fruit set percentage was 92.3% observed 20 days after pollination.
Example 2 (pollen sample two):
1) subpackaging the picked kiwi fruit pollen, sealing in plastic bottles with the height of 3cm and the thickness of 3mm, 10 g/bottle, totaling 200 bottles, placing in an ultrasonic cleaner, performing ice-water bath with the power of 40W, and treating for 10 minutes;
2) immediately feeding the pollen after ultrasonic treatment into the container60Co-gamma ray irradiation is carried out in an irradiation cabin, the irradiation dose is 600Gry, and the temperature is 28 ℃;
3) storing the pollen after irradiation at-20 deg.C;
4) taking out pollen before pollination, and standing at room temperature for 3 hours;
5) fully mixing pollen and lycopodium clavatum powder according to the mass ratio of 1:0.7, and pollinating 12,734 female flowers on 50 kiwi trees;
6) pollen viability was determined by pollen germination method with control pollen sample viability of 57.2%, irradiation sample viability of 27.3%, and PSA detection negative.
7) Observed 20 days after pollination, the fruit setting rate is 94.6 percent.
Example 3 (pollen sample three):
1) subpackaging and sealing the picked kiwi fruit pollen in plastic bottles with the height of 3cm and the thickness of 3mm, 10 g/bottle, totaling 200 bottles, placing in an ultrasonic cleaner, carrying out ice-water bath with the power of 60W, and treating for 3 minutes;
2) immediately feeding the pollen after ultrasonic treatment into the container60Co-gamma ray irradiation is carried out in an irradiation cabin, the irradiation dose is 400Gry, and the temperature is 20 ℃;
3) storing the pollen after irradiation at-20 deg.C;
4) taking out pollen before pollination, and standing at room temperature for 3 hours;
5) fully mixing pollen and lycopodium clavatum powder according to the mass ratio of 1:0.7, and pollinating 14,121 female flowers on 50 kiwi trees;
6) pollen viability was determined by pollen germination method, with control pollen sample viability of 47.9%, irradiated pollen sample viability of 39.4%, and PSA detection negative.
7) Observed 20 days after pollination, the fruit setting rate is 96.3 percent.
At present, no report is found on related technologies for keeping kiwi fruit pollen alive and killing PSA.
The above description is only a preferred embodiment of the present invention, and is not intended to limit the invention in any way, and any simple modification, equivalent change and modification made to the above embodiment according to the technical spirit of the present invention are within the scope of the technical solution of the present invention without departing from the technical solution of the present invention.
Claims (1)
1. A method for killing ulcer disease pathogenic bacteria PSA after kiwi pollen is picked is characterized in that: the method comprises the following steps:
1) subpackaging the picked kiwi fruit pollen, sealing in plastic bottles with the height of 3cm and the thickness of 3mm, 10 g/bottle, totaling 200 bottles, placing in an ultrasonic cleaner, performing ice-water bath with the power of 40W, and treating for 10 minutes;
2) immediately feeding the pollen after ultrasonic treatment into the container60Co-gamma ray irradiation is carried out in an irradiation cabin, the irradiation dose is 600Gry, and the temperature is 28 ℃;
3) storing the pollen after irradiation at-20 deg.C;
4) taking out pollen before pollination, and standing at room temperature for 3 hours;
5) fully mixing pollen and lycopodium clavatum powder according to the mass ratio of 1:0.7, and pollinating 12,734 female flowers on 50 kiwi trees;
6) pollen viability is measured by a pollen germination method, the control pollen sample viability is 57.2%, the irradiation sample viability is 27.3%, and PSA detection is negative;
7) observed 20 days after pollination, the fruit setting rate is 94.6 percent.
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| CN111514320B (en) * | 2020-03-23 | 2021-08-10 | 贵阳学院 | Method for killing ulcer disease pathogenic bacteria PSA in kiwi fruit pollen |
| CN111420084B (en) * | 2020-03-23 | 2021-09-24 | 贵阳学院 | Method for killing pathogenic bacteria (PSA) of kiwi fruit pollen canker in vacuum by using low-temperature microwaves |
| CN112294982A (en) * | 2020-09-21 | 2021-02-02 | 中国科学院武汉植物园 | Cobalt radiation sterilization method for kiwi fruit pollen with canker pathogen |
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| US20150283276A1 (en) * | 2014-04-08 | 2015-10-08 | The United States Of America, As Represented By The Secretary Of Agriculture | Method for Controlling Fungal Plant Pathogens Using a Combination of UV Radiation Followed by Antagonist Application and Dark Period |
| CN104872111B (en) * | 2015-05-22 | 2017-01-25 | 桂林嘉佰农农业科技有限公司 | Storing method of pollens subjected to soft x ray irradiation |
| CN106035060B (en) * | 2016-05-30 | 2018-10-26 | 陕西师范大学 | A kind of method of Psa in killing Kiwi fruit pollen |
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