CN102415370B

CN102415370B - Ultralow temperature preservation method for capsicum pollens

Info

Publication number: CN102415370B
Application number: CN 201110237274
Authority: CN
Inventors: 白占兵; 李雪峰; 戴雄泽; 马艳青; 陈文超; 张竹青; 周书栋; 邹学校
Original assignee: INST OF VEGETABLES HUNAN PROV
Current assignee: INST OF VEGETABLES HUNAN PROV
Priority date: 2011-08-18
Filing date: 2011-08-18
Publication date: 2013-05-08
Anticipated expiration: 2031-08-18
Also published as: CN102415370A

Abstract

The invention provides a preservation method for paternal pollens in the capsicum hybrid production. In the capsicum hybrid production, male parents and female parents are planted in a time staggering and allopatry mode, and the pollens of the male parents, which are concentratedly stored in liquid nitrogen, are transported to female parent planting sites for the usage during pollination. According to the invention, the flowering asynchronism of the male parents and the female parents in the capsicum hybrid production is thoroughly solved, so the simultaneous and sympatric appearance of capsicum at the seed production male parents and the seed production female parents is avoided, and the loss of the parent of the male parents can be effectively prevented; and superfluous pollens in the seed production can be stored, so the stored pollens can be used to cover the shortage when the amount of the pollens of the male parents is not enough.

Description

Capsicum pollens ultralow temperature is preserved

Affiliated technical field

The present invention is the store method of a cover capsicum pollens, relates to the production of hybrid seeds of capsicum business, specifically the application in a kind of capsicum pollens cryopreservation method and the business production of hybrid seeds.

Background technology

In capsicum cross breeding seed process often the Parent flowering asynchronism can appear; the paternal pollen surplus causes unnecessary waste; the impact that is subject to natural cause causes pollen to lack; affect the output of seed; be planted in easily stolenly together through the kind that meticulously selects for many years Parent when the production of hybrid seeds, these problems are perplexing whole world breeding man and seeds company always.If male parent is long-term with the form of pollen, safety is preserved; can offer at any time production of hybrid seeds family; can thoroughly solve the problem of the Parent flowering asynchronism of the production of hybrid seeds; the intellectual property of protection kind; from land rent and the whole pollen of the comparatively cheap local plantation male parent collection of labour; can greatly reduce breeding cost, so the low temperature of capsicum pollens is preserved significant.

Summary of the invention

In order to overcome in capsicum cross breeding seed process often the Parent flowering asynchronism can appear; pollen is superfluous or lack; the parent such as easily loses at the series of problems; the invention provides the capsicum store method of a cover safety; the pollen of preserving can use at any time, can thoroughly solve the problem of the flowering asynchronism that exists in the capsicum cross breeding seed, and male parent can shift to an earlier date half a year or more early be planted in more hidden place; the male parent of new varieties is difficult for losing like this, has closely protected new varieties.

The technical solution adopted for the present invention to solve the technical problems is: its technology comprises following step: (1) capsicum full-bloom stage gathers well-developed pollen; (2) pollen is dried to suitable water content 8-13%, is encapsulated in the cryovial of 2.0 milliliters; (3) through-20 ℃ of precooling treatment 0.5 hour or directly put into the liquid nitrogen container that liquid nitrogen is housed and preserve; (4) will fill polliniferous liquid nitrogen container and be transported to planting of maternal ground; (5) as required, take out at any time cryovial, naturally thaw, make the pollen recovery; (6) have blodynamic pollen and be used for business production of hybrid seeds production.

The invention has the beneficial effects as follows: can thoroughly solve the flowering asynchronism problem in the capsicum production of hybrid seeds; in the production of hybrid seeds, male parent is given production of hybrid seeds family with the form of pollen; male parent can be planted in any can be planted in the place that is fit to the growth capsicum; the traversing space-time production of hybrid seeds; plant in advance male parent half a year; also can arrange female parent according to the amount of male parent, protect male parent to lose in the production of hybrid seeds.

Embodiment

(1) store method: the florescence gathers capsicum, take out flower pesticide, flower pesticide is dispersed on glazed paper equably, and in the environment of drying, pollen exposes from flower pesticide, sieve after impurity elimination with 120 mesh sieve, pollen after sieving is wrapped with glazed paper, put in the airtight canopy that quicklime is housed, making pollen water content is 8-13%, then dry pollen is packed in 2.0 milliliters of cryovials, through-20 ℃ of precooling treatment 0.5 hour or directly put into liquid nitrogen and preserve; Take out the test tube seal pollen up for safekeeping when female parent need to be pollinated, Indoor Natural give birth to temperature to test tube internal and external temperature one to, make the pollen recovery; In time regain residue pollen after pollination finishes, tubulature is preserved among dropping into liquid nitrogen again, for future use.

(2) case study on implementation and analysis

Embodiment 1

The relation of different pretreatments, defreezing method and Pollen Activity

In condition at one's leisure, do pretreatment with-20 ℃ before pollen storage better, also can not do pretreatment and directly will be equipped with in the cryovial input liquid nitrogen of capsicum pollens; Directly taking out to thaw to thaw with other method at Indoor Natural when thawing does not have significant difference, therefore selects directly and thaws at Indoor Natural.

Table 1: the storage pollen of one month is in the relation (experimental period: 2009.2.18) of different pretreatments, different defreezing method and Pollen Activity in liquid nitrogen

Embodiment 2

Table 2: the bimestrial pollen of storage is in the relation (experimental period: 2009.3.23) of different pretreatments, different defreezing method and Pollen Activity in liquid nitrogen

Annotate: adopt TTC Determination Staining Pollen Activity in table 1, table 2, the slice, thin piece of three repetitions is done in every processing, and each slice, thin piece is observed 5 visuals field, calculates average result

Embodiment 3

Table 3 is preserved the pollen of half a year in the relation (experimental period: 2009.6.) of different pretreatments, different defreezing method and Pollen Activity in liquid nitrogen

Illustrate: A: represent 4 ℃ of precoolings; B: represent-20 ℃ of precoolings; C: represent in the liquid nitrogen that directly drops into-196 ℃; 2009 because running into high temperature, so the pollination ripening rate is lower; The pollen of above table 1,2,3 storages were adopted in Sanya, Hainan on January 15th, 2009, and January 17 is to Changsha, in the test tube of 2.0 milliliters of packing into January 18, did to throw in liquid nitrogen after different processing; 2009.6.16-6.19 a day pollen thawing mode was put 30 minutes for-20 ℃, 6.20-6.30 pollen solves the mode of freezing direct nature that takes out from liquid nitrogen and thaws.Therefore ,-20 ℃ of precoolings or directly drop into better during input for instant noodles are selected direct input, had not only facilitated Pollen Activity but also high with the nature freezing process when thawing.

Embodiment 4

Capsicum pollens water content and its vigor relation

Fine day is placed in the pollen that sieves under the closed environment of depositing quicklime, makes pollen water content reach 8-13%, and drop in liquid nitrogen and can preserve safely, at the pollen of cloudy undried, the pollen variable color, vigor descends obviously; Table 4 result shows: the pollen of fine day can direct plunge into Liquid Nitrogen among, cloudy pollen just can be put into liquid nitrogen after must drying and preserve, before the pollen storage that fine day gathers, drying effect is best, also can direct plunge into Liquid Nitrogen in.

Table 4 capsicum pollens water content and its vigor relation (in December, 2009 Sanya)

Illustrate: pollination combination LS20A * J0721, pollen water content is to be measured by water analysis instrument Ao Haosi MB23;

Embodiment 5

Combination 14A * J0721, pollen picked up from Sanya, Hainan on December 12nd, 2009, and from June 12nd, 2010 to June 21,20 flowers are pollinated every day;

Result such as following table: June 15,16 days, three days on the 18th all arrange contrast, all there is no difference with comparison film by statistics;

Table 5 is preserved fruit-setting rate, the solid number of the pollen of half a year

Embodiment 6

Preservation half a year, 1 year pollen is used for the production of hybrid seeds, the rich capsicum of production kind No. five, pollen respectively at adopt in the Sanya in December, 2009, adopt in Changsha in June, 2010, it is Nan Bin farm, Sanya that the place is implemented in the production of hybrid seeds, in September, 2010-2011 year March production of hybrid seeds time, each 333.3 square metres of production of hybrid seeds areas contrast 333.3 square metres.

The different storages of table 6 capsicum pollens 1 year, half a year, fresh hybrid seed yield contrast

The pollen storage time	1 year	Half a year	Powder CK now picks flowers
				Output (KG)	29.5	28	28.6

Claims

1. the ultralow temperature of capsicum pollens is preserved, it is characterized in that: the capsicum florescence gathers well-developed pollen, pollen is dried to water content 8-13%, is encapsulated in cryovial, be equipped with in the liquid nitrogen container of liquid nitrogen through-20 ℃ of precooling treatment 0.5 hour or direct the input, preserve a period of time, be transported to planting of maternal ground, as required, take out from liquid nitrogen container at any time and seal the pollen cryovial up for safekeeping, room temperature is slowly thawed, and makes the pollen recovery.

2. capsicum pollens ultralow temperature according to claim 1 is preserved, it is characterized in that: be used for solving the capsicum crossbreed with liquid nitrogen storage capsicum pollens and produce the flowering asynchronism problem: be about to pollen storage unnecessary in the production of hybrid seeds in liquid nitrogen, or when in the production of hybrid seeds, the paternal pollen amount is inadequate, cover the shortage with the pollen of preserving in liquid nitrogen.

3. capsicum pollens ultralow temperature according to claim 1 is preserved, it is characterized in that: in capsicum production of hybrid seeds process, Parent is staggered the time plant or Planting in the different location or the also Planting in the different location of staggering the time, the pollen of male parent is concentrated be housed in liquid nitrogen, be transported to planting of maternal ground use when pollination.