CN103392450A - Method for collecting rice anther - Google Patents
Method for collecting rice anther Download PDFInfo
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- CN103392450A CN103392450A CN2013103427377A CN201310342737A CN103392450A CN 103392450 A CN103392450 A CN 103392450A CN 2013103427377 A CN2013103427377 A CN 2013103427377A CN 201310342737 A CN201310342737 A CN 201310342737A CN 103392450 A CN103392450 A CN 103392450A
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Abstract
The invention discloses a method for collecting rice anther. The method includes the following steps: first, taking rice ears, sterilizing the rice ears and then removing the top ends of glumes, and second, placing the ear portions into a collection bottle in a suspended mode with stalks of rice buds facing upwards and the ear portions of the rice buds facing downwards, and vibrating the collection bottle to collect and obtain the anther. The method for collecting the rice anther is simple in operation, the speed of collecting the rice anther is higher by more than one time than that in a conventional collection method, contact pollution will not happen in the process of inoculation, mechanical damage to the anther can be avoided, continuous production can be achieved, and any environmental pollution will not be produced.
Description
Technical field
The present invention relates to the plant anther collection method, relate in particular to a kind of collection method of Rice Anther.
Background technology
Anther culture is to pass through plant tissue culture technique, by the flower pesticide of growing to certain phase, by aseptic technique, be seeded on synthetic medium, to change the development program of pollen grain in flower pesticide, induce its differentiation, and carry out continuously mitosis, form cell mass, and then form a parenchyma-callus without differentiation, or be divided into embryoid, lure that subsequently Calli Differentiation becomes complete plant into.
In China, the Rice Anther culture technique is mainly used in three aspects: the first, and will spend training technology (anther culture technique) to combine with the breeding of conventional variety intermolecular hybrid, carry out the cultivation of new varieties, this aspect has obtained significant achievement.The second, will spend the training technology for Xian, the breeding of round-grained rice inter-subspecies hybrid, the kind that this respect is bred as is many.The 3rd, will spend the training technology to purificate and rejuvenate for hybrid rice, with flower pesticide purify more new sterile line, maintainer, its yield increased volume increase 10%.At present, the plant that flower pesticide obtains is set up for the theoretical foundation of breeding, paddy rice has become the crop that can utilize the anther culture technique rapid breeding.
In the anther culture of paddy rice, comparatively suitable to inducing haplobiont with the keep to the side pollen of phase of monokaryon, cultivate this period to flower pesticide, induce efficiency high, and whether pollen is in the monokaryon phase of keeping to the side, can judges and identify by some morphological indexs, as pulvinus distance, rice glume color, stamen length etc.
Getting flower pesticide is that the first step that Rice Anther is cultivated is also very important step, the raw material that flower pesticide is cultivated as tissue, when getting flower pesticide, should avoid flower pesticide is damaged, in order to avoid efficiency is induced in impact, also need simultaneously to reduce the probability that pollutes as far as possible, should consider in addition the convenience of experimental implementation.At present, when getting flower pesticide, usually first spike of rice is carried out disinfection, after sterilization, in superclean bench, by eye scissors, the clever shell of spike of rice is ripped, take out the flower pesticide postposition and with ophthalmic tweezers, it is inoculated in medium and cultivates.There is following problem in the method: (1) complex operation and difficulty are high, and labour intensity is large, and be time-consuming, is difficult for scale and carries out production operation; (2) in operating process, the utensils such as ophthalmic tweezers, eye scissors directly contact flower pesticide or medium, easily cause contact stain, introduce miscellaneous bacteria, pollute medium, simultaneously also very easily to the flower pesticide injury, affect callus induction rate.
Summary of the invention
For overcoming above-mentioned defect, the invention provides a kind of collection method of Rice Anther, easy to operate, collection efficiency is high and reduced anther cultural pollution probability.
A kind of collection method of Rice Anther comprises:
Get spike of rice, after sterilization, remove the top of clever shell; By rice bud stem stalk up, fringe section makes described spike of rice be suspended in receiving flask down, and vibration is collected and obtained described flower pesticide.
While removing the top of clever shell, can adopt the method for wiping out or excising.
Because the flower pesticide of paddy rice is wrapped up by clever shell, the top of clever shell to be removed, the release that is flower pesticide provides passage, and by vibration, flower pesticide can fall under gravity in receiving flask.
Sword-like leave pulvinus to the 2 leaves that rice bud is extracted out, when light green color appears in clever shell from described rice bud, taking out spike of rice.At this moment, the pollen of paddy rice is in monokaryon and keeps to the side the phase, and the flower pesticide of the rice of fetching water this period carries out cultured in vitro, and callus induction rate is high.
Before sterilization, described spike of rice was placed 4～6 days in 4～6 ℃.Suitable low temperature pretreatment can slow down degeneration and the brown stain of anther wall, impels the flower pesticide dedifferentiation to form callus.
The method of described sterilization comprises: described spike of rice, with after 70% alcohol disinfecting 30s～1min, is placed in spike of rice to 1% clorox and soaks 10～15min.
While removing the top of clever shell, the part that clever shell is removed accounts for 1/3～1/2 of described clever shell.If the part of removing is less than 1/3, clever shell opener is little, and flower pesticide is difficult for being vibrated, and affects collecting effect; If the clever shell of removing, greater than 1/2, easily makes flower pesticide be subject to mechanical damage.
Distance between at the bottom of described fringe section and receiving flask bottle is 2～6cm, is preferably 3～5cm.Distance is too small, and the collected flower pesticide distribution of vibrating is less, causes the flower pesticide skewness, and local Anther density is large; And distance is excessive, and flower pesticide is easily oscillated on the receiving flask wall, affects collecting effect.
In described receiving flask, be loaded with agar filter paper.Flower pesticide is collected on agar filter paper, and agar filter paper adheres to flower pesticide, can directly agar filter paper be shifted and carry out cultured in vitro, and is easy to operate.
It is two-layer that described receiving flask is divided into up and down, and upper strata is provided with the fixture for fixing described spike of rice, and lower floor is loaded with described agar filter paper.Receiving flask is set to two-layer, has facilitated spike of rice is fixed, and due to flower pesticide, dropped into the lower floor of receiving flask, after the flower pesticide collection is complete, two-layer the taking apart in up and down can be shifted the lower floor that fills flower pesticide to operate, and reduces and pollutes probability.
The frequency of described vibration is 0.1～10 hertz, is preferably 3～10 hertz.The falling speed that the frequency of vibrating by control can be regulated flower pesticide, but frequency of oscillation is too high and too low all improper, if frequency of oscillation is too low, flower pesticide is difficult for falling, and affects the collection efficiency of flower pesticide, and the too high flower pesticide that easily causes of frequency of oscillation adheres to the receiving flask inwall, affect collecting effect, in addition, frequency of oscillation excessive also easily by on spike of rice impurity shake, the effect that impact is collected.
The time of described vibration is 1～30s, is preferably 8～15s.Suitable duration of oscillation can guarantee to collect the flower pesticide of q.s, also can avoid simultaneously duration of oscillation long and cause falling into of impurity on spike of rice.
Compared with prior art, beneficial effect of the present invention is:
(1) with tradition with tweezer, cut and peel off the method for getting flower pesticide and compare, the present invention can carry out the collection of flower pesticide by vibration, more convenient to operate, gathering speed also improves greatly than it, but the continuous large-scale collection does not produce any environmental pollution.
(2) than conventional method, compare, the present invention is in the flower pesticide collection process, operation tool such as tweezers etc. directly do not contact flower pesticide, can not cause mechanical damage and pollution to flower pesticide, during the inoculation cultured in vitro, do not need directly to contact flower pesticide and culture matrix yet, avoided contact stain, reduced the pollution probability.
(3) the present invention, in the flower pesticide collection process, by regulating frequency of oscillation, controls the flower pesticide falling speed, the collection of flower pesticide is more flexible, without with tweezers from picking flower pesticide clever shell, help relieving eye strain and operating personnel's continuous firing state, operate easier.
The accompanying drawing explanation
Fig. 1 is the schematic diagram that embodiment 1～2 receiving flask is collected flower pesticide.
Fig. 2 is the callus that embodiment 2 anther culture form.
Embodiment
Below in conjunction with specific embodiment, the present invention is further explained.
Fig. 1 has shown that embodiment 1～2 receiving flask collects the schematic diagram of flower pesticide, and visible in figure, receiving flask is cylindric, is divided into top cylinder, doffing is two-layer, and top cylinder and doffing can be threaded connection.The top of top cylinder 2 is along being fixed with a support bar 1, and support bar 1 is axial vertical with receiving flask, and the middle part of support bar 1 is with a clip 8, and for the stem stalk of clamping spike of rice 6, the bottom of doffing 3 is loaded with agar filter paper 4.While collecting flower pesticide, whole receiving flask is placed on oscillator 5, and the vibration of oscillator 5 shakes flower pesticide fall.
The method of Rice Anther is collected in vibration in receiving flask, comprises the following steps:
(1) sword-like leave pulvinus to the 2 leaves extracted out of rice bud, get fringe in field when light green color appears in spike of rice grain husk shell, by spike of rice in 4 degrees centigrade of low temperature pretreatment 5 days.
(2) spike of rice after processing is placed in the sterile tissue blake bottle, and the alcohol with 70% carries out surface sterilization 1min, removes alcohol, in single aseptic superclean bench, with 1% clorox sterilization 15min, removes clorox, with aseptic water washing 5 times.
(3) spike of rice after sterilizing cuts off the part more than 1/3 on clever shell top with scissors in single aseptic super-clean bench, in Fig. 1, has shown complete clever shell 6, removes the clever shell 7 on top.
(4) the receiving flask bottom is set off to aseptic agar filter paper under aseptic condition, by the fringe section that cuts off clever shell down, the stem stalk is put into receiving flask up, tightens two layers (being top cylinder, doffing) in receiving flask up and down, and the stem stalk is clipped, the distance between at the bottom of the top of fringe section and receiving flask bottle is 3cm.
(5) receiving flask is put into vibration collection on oscillator, and frequency of oscillation is adjusted in 3 hertz, vibration 15s, and flower pesticide is collected in receiving flask substantially.
(6) collect after completing and turn on receiving flask in aseptic super-clean bench, take out the agar filter paper of bottle bottom, will transfer in culture dish with the agar filter paper of flower pesticide, be placed in 25 ℃ of incubators and secretly be cultured to and induce callus.
Can collect 24 bottles of flower pesticide in two hours, and every bottle of flower pesticide >=150 piece, the flower pesticide of collection is cultivated, after secretly cultivating 35 days, the statistics callus induction rate, its inductivity is 10.16%, and does not produce any pollution.
The method of Rice Anther is collected in vibration in receiving flask, comprises the following steps:
(1) rice bud extract out sword-like leave pulvinus to the 2 leaf the time, when light green color appears in spike of rice grain husk shell, in field, get fringe, by spike of rice in 5 degrees centigrade of low temperature pretreatment 6 days.
(2) spike of rice after processing is placed in the sterile tissue blake bottle, and the alcohol with 70% carries out surface sterilization 1min, removes alcohol, in single aseptic superclean bench, with 1% clorox sterilization 15min, removes clorox, with aseptic water washing 5 times.
(3) after the sterilization of the spike of rice after processing in the clean platform of double aseptic behaviour with scissors partly cutting off 1/2 or more clever shell top.
(4) in the clean platform of aseptic behaviour, the receiving flask bottom is set off to aseptic agar filter paper, by the fringe section that cuts off clever shell down, the stem stalk is put into receiving flask up, and the stem stalk is clipped, tighten two layers (being top cylinder, doffing) in receiving flask up and down, between at the bottom of the top of spike of rice and receiving flask bottle apart from being 5cm.
(5) receiving flask is put into vibration collection on oscillator, and frequency of oscillation is adjusted in 10 hertz, and duration of oscillation is 8s.
(6) collect after completing and turn on receiving flask in the clean platform of aseptic behaviour, take out the agar filter paper of bottle bottom, flower pesticide is transferred in culture dish, be placed in 25 ℃ of incubators and secretly be cultured to and induce callus.
Can collect 26 bottles of flower pesticide in two hours, and every bottle of flower pesticide >=150 piece, after secretly cultivating 25 days, the statistics callus induction rate, its inductivity is 10.35%, and does not produce any pollution, Fig. 2 shows the callus that the present embodiment anther culture forms.
Comparative Examples
(1) sword-like leave pulvinus to the 2 leaves extracted out of rice bud, get spike of rice in field when light green color appears in spike of rice grain husk shell, by spike of rice in 5 degrees centigrade of low temperature pretreatment 6 days.
(2) spike of rice after processing is placed in the sterile tissue blake bottle, and the alcohol with 70% carries out surface sterilization 1min, removes alcohol, in single aseptic superclean bench, with 1% clorox sterilization 15min, removes clorox, with aseptic water washing 5 times.
(3) while getting flower pesticide, adopt conventional method, first by eye scissors, the clever shell of spike of rice is ripped, taken out flower pesticide and be placed in sterile petri dish, after flower pesticide has some, with ophthalmic tweezers, it is inoculated in medium, is placed in 25 ℃ of incubators and secretly is cultured to and induces callus.
By statistics, 1200 pieces of flower pesticide are got in people's operation, approximately 2 hours, these 1200 pieces of flower pesticide are inoculated in medium, 150 pieces of each culture dish inoculations, totally 8 culture dishes, cultivate after 35 days, the statistics callus induction rate, its inductivity is 9.95%, and finds that there is two wares generation pollutions.
Claims (10)
1. the collection method of a Rice Anther comprises:
Get spike of rice, after sterilization, remove the top of clever shell; By rice bud stem stalk up, fringe section makes described fringe section be suspended in receiving flask down, and vibration is collected and obtained described flower pesticide.
2. collection method as claimed in claim 1, is characterized in that, sword-like leave pulvinus to the 2 leaves that rice bud is extracted out, when light green color appears in clever shell from described rice bud, taking out spike of rice.
3. collection method as claimed in claim 1, is characterized in that, before sterilization, described spike of rice placed 4～6 days in 4～6 ℃.
4. collection method as claimed in claim 1, is characterized in that, the method for described sterilization comprises: described spike of rice, with after 70% alcohol disinfecting 30s～1min, is placed in spike of rice to 1% clorox and soaks 10～15min.
5. collection method as claimed in claim 1, is characterized in that, the part that clever shell is removed accounts for 1/3～1/2 of described clever shell.
6. collection method as claimed in claim 1, is characterized in that, the distance between at the bottom of described fringe section and receiving flask bottle is 2～6cm.
7. collection method as claimed in claim 1, is characterized in that, in described receiving flask, is loaded with agar filter paper.
8. collection method as claimed in claim 7, is characterized in that, it is two-layer that described receiving flask is divided into up and down, and upper strata is provided with the fixture for fixing described spike of rice, and lower floor is loaded with described agar filter paper.
9. collection method as claimed in claim 1, is characterized in that, the frequency of described vibration is 0.1～10 hertz.
10. collection method as claimed in claim 1, is characterized in that, the time of described vibration is 1～30s.
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| CN201310342737.7A CN103392450B (en) | 2013-08-07 | 2013-08-07 | A kind of collection method of Rice Anther |
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| CN201310342737.7A CN103392450B (en) | 2013-08-07 | 2013-08-07 | A kind of collection method of Rice Anther |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106119076A (en) * | 2016-06-29 | 2016-11-16 | 无锡南理工科技发展有限公司 | A kind of crops training spore suspension preparation facilities and method |
Citations (5)
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| KR20080050759A (en) * | 2006-12-04 | 2008-06-10 | 임장혁 | Flower pine pick catch system |
| CN102217464A (en) * | 2011-05-03 | 2011-10-19 | 西安建筑科技大学 | Combined cyclone separation/collection device and method for separating and collecting pollen |
| CN102793728A (en) * | 2012-08-24 | 2012-11-28 | 安徽哈博药业有限公司 | High-frequency oscillation wall-broken ganoderma lucidum spore powder |
| CN102907314A (en) * | 2012-11-09 | 2013-02-06 | 韦炳辉 | Selection and breeding method for early rice variety |
| CN202857345U (en) * | 2012-10-09 | 2013-04-10 | 谭玉丰 | Anther collector |
-
2013
- 2013-08-07 CN CN201310342737.7A patent/CN103392450B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20080050759A (en) * | 2006-12-04 | 2008-06-10 | 임장혁 | Flower pine pick catch system |
| CN102217464A (en) * | 2011-05-03 | 2011-10-19 | 西安建筑科技大学 | Combined cyclone separation/collection device and method for separating and collecting pollen |
| CN102793728A (en) * | 2012-08-24 | 2012-11-28 | 安徽哈博药业有限公司 | High-frequency oscillation wall-broken ganoderma lucidum spore powder |
| CN202857345U (en) * | 2012-10-09 | 2013-04-10 | 谭玉丰 | Anther collector |
| CN102907314A (en) * | 2012-11-09 | 2013-02-06 | 韦炳辉 | Selection and breeding method for early rice variety |
Non-Patent Citations (1)
| Title |
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| 王辉等: "水稻花药愈伤组织诱导的多因子正交试验研究", 《广西植物》, vol. 31, no. 4, 31 July 2011 (2011-07-31), pages 545 - 549 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106119076A (en) * | 2016-06-29 | 2016-11-16 | 无锡南理工科技发展有限公司 | A kind of crops training spore suspension preparation facilities and method |
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Inventor after: Deng Li Inventor after: Wu Zhongming Inventor after: Lv Xiaoliang Inventor after: Zhang Yuexiao Inventor after: Wang Qing Inventor after: Feng Duhua Inventor after: Fu Junjie Inventor after: Shu Qingyao Inventor after: Cui Hairui Inventor after: Huang Jianzhong Inventor after: Sun Xiaoqiang Inventor before: Feng Duhua Inventor before: Lv Xiaoliang Inventor before: Wang Qing Inventor before: Deng Li Inventor before: Fu Junjie Inventor before: Shu Qingyao Inventor before: Cui Hairui Inventor before: Huang Jianzhong Inventor before: Sun Xiaoqiang Inventor before: Wu Zhongming |
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Effective date of registration: 20150917 Address after: 214104 modern agricultural exhibition garden, anzhen Town, Xishan District, Jiangsu, Wuxi Applicant after: WUXI HUPPER SEEDS BIOLOGICAL AGRICULTURE CO., LTD. Applicant after: Wuxi Qiushi Biological Agricultural Co., Ltd. Address before: East Lake Tong East Avenue Xishan District 214196 in Jiangsu province Wuxi city Wuxi high tech agricultural demonstration garden Applicant before: Wuxi Qiushi Biological Agricultural Co., Ltd. |
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