CN103392450B - A kind of collection method of Rice Anther - Google Patents
A kind of collection method of Rice Anther Download PDFInfo
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- CN103392450B CN103392450B CN201310342737.7A CN201310342737A CN103392450B CN 103392450 B CN103392450 B CN 103392450B CN 201310342737 A CN201310342737 A CN 201310342737A CN 103392450 B CN103392450 B CN 103392450B
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Abstract
The invention discloses a kind of collection method of Rice Anther, comprising: get spike of rice, after sterilization, remove the top of clever shell; By rice bud stem stalk upward, fringe portion makes described fringe portion be suspended in receiving flask down, and vibration is collected and obtained described flower pesticide.The collection method of Rice Anther of the present invention, easy to operate, more than the gathering speed of Rice Anther is fast again compared with Regular method, does not produce contact stain during inoculation, mechanical damage is not caused to flower pesticide, can continuous seepage, do not produce any environmental pollution.<pb pnum="1" />
Description
Technical field
The present invention relates to plant anther collection method, particularly relate to a kind of collection method of Rice Anther.
Background technology
Anther culture passes through plant tissue culture technique, flower pesticide to certain phase will be grown, by aseptic technique, be seeded on synthetic medium, to change the development program of pollen grain in flower pesticide, induce it to break up, and carry out mitosis continuously, form cell mass, and then form a parenchyma-callus without differentiation, or be divided into embryoid, lure the plant that Calli Differentiation becomes complete subsequently into.
In China, Rice anther culture technology is mainly used in three aspects: the first, and flower training technology (anther culture technique) combined with the breeding of conventional variety intermolecular hybrid, carry out the cultivation of new varieties, this aspect has achieved significant achievement.The second, flower training technology is used for Xian, the breeding of Via-back puncture intermolecular hybrid, the variety protection that this respect is bred as is many.3rd, flower training technology is used for hybrid rice purification and rejuvenation, purifies more new sterile line, maintainer with flower pesticide, its yield increased volume increase 10%.At present, the theoretical foundation that the plant obtained by flower pesticide is used for breeding is set up, and paddy rice has become the crop that can utilize anther culture technique rapid breeding.
In the anther culture of paddy rice, comparatively suitable to induction haplobiont with the pollen of mid-late uninucleate stage, this period cultivates flower pesticide, induced efficiency is high, and whether pollen is in mid-late uninucleate stage by some morphological indexs is undertaken judging and identifying, as pulvinus distance, rice glume color, stamen length etc.
To get flower pesticide be the first step of Rice anther culture is also very important step, flower pesticide is as the raw material of tissue cultures, when getting flower pesticide, should avoid damaging flower pesticide, in order to avoid affect induced efficiency, also need the probability of decreasing pollution as far as possible simultaneously, the convenience of experimental implementation should be considered in addition.At present, when getting flower pesticide, usually first being carried out disinfection by spike of rice, after sterilization, in superclean bench, being ripped by the clever shell of eye scissors by spike of rice, the rearmounted ophthalmic tweezers of taking-up flower pesticide is inoculated in medium to be cultivated.There is following problem in the method: (1) complex operation and difficulty is high, and labour intensity is large, and time-consuming, not easily production operation is carried out in scale; (2) in operation, the utensil such as ophthalmic tweezers, eye scissors directly contacts flower pesticide or medium, easily causes contact stain, introduces miscellaneous bacteria, pollutes medium, also very easily causes damage to flower pesticide simultaneously, affects 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 reduce anther cultural pollution probability.
A collection method for Rice Anther, comprising:
Get spike of rice, after sterilization, remove the top of clever shell; By rice bud stem stalk upward, fringe portion makes described spike of rice be suspended in receiving flask down, and vibration is collected and obtained described flower pesticide.
When removing the top of clever shell, the method for wiping out or excising can be adopted.
Because the flower pesticide of paddy rice is wrapped up by clever shell, removed on the top of clever shell, the release being flower pesticide provides passage, and by vibration, flower pesticide can fall under gravity in receiving flask.
The sword-like leave pulvinus that rice bud is extracted out, to the 2nd leaf, takes out spike of rice when light green color appears in clever shell from described rice bud.Now, the pollen of paddy rice is in mid-late uninucleate stage, and the flower pesticide of rice of fetching water this period carries out cultured in vitro, and callus induction rate is high.
Before sterilization, described spike of rice is placed 4 ~ 6 days in 4 ~ 6 DEG C.Suitable Cold pretreatment can slow down degeneration and the brown stain of anther wall, impels flower pesticide dedifferentiation to form callus.
The method of described sterilization comprises: by described spike of rice with after 70% alcohol disinfecting 30s ~ 1min, and clorox spike of rice being placed in 1% soaks 10 ~ 15min.
When 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 removed is less than 1/3, then clever shell opener is little, and flower pesticide is not easily vibrated, and affects collecting effect; If the clever shell removed is greater than 1/2, then flower pesticide is easily made to be subject to mechanical damage.
Distance between at the bottom of described fringe portion and receiving flask bottle is 2 ~ 6cm, is preferably 3 ~ 5cm.Apart from too small, the flower pesticide distribution collected by vibration is less, causes flower pesticide skewness, and local Anther density is large; And apart from excessive, flower pesticide is easily oscillated on receiving flask wall, affect collecting effect.
Agar filter paper is loaded with in described receiving flask.Flower pesticide is collected on agar filter paper, and flower pesticide adheres to by agar filter paper, directly the transfer of agar filter paper can be carried out cultured in vitro, easy to operate.
Described receiving flask is divided into two-layer 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, facilitates and spike of rice is fixed, and drop into the lower floor of receiving flask due to flower pesticide, after flower pesticide is collected, can take apart two-layer up and down, the lower floor's transfer filling flower pesticide is operated, decreasing pollution probability.
The frequency of described vibration is 0.1 ~ 10 hertz, is preferably 3 ~ 10 hertz.The falling speed of flower pesticide can be regulated by the frequency controlling vibration, but frequency of oscillation is too high and too low all improper, if frequency of oscillation is too low, then flower pesticide not easily falls, and affect the collection efficiency of flower pesticide, frequency of oscillation is too high, easily causes flower pesticide to adhere to receiving flask inwall, affect collecting effect, in addition, frequency of oscillation excessive also easy by spike of rice impurity shake, impact collect effect.
The time of described vibration is 1 ~ 30s, is preferably 8 ~ 15s.Suitable duration of oscillation can ensure the flower pesticide collecting q.s, duration of oscillation also can be avoided long simultaneously and cause falling into of impurity on spike of rice.
Compared with prior art, beneficial effect of the present invention is:
(1) with tradition tweezer, cut compared with the method peeling off and get flower pesticide, the present invention can carry out the collection of flower pesticide by vibration, more convenient to operate, and also comparatively it improves gathering speed greatly, can gather on a large scale continuously, not produce any environmental pollution.
(2) comparatively conventional method is compared, the present invention is in flower pesticide collection process, operation tool such as tweezers etc. directly do not contact flower pesticide, mechanical damage and pollution can not be caused to flower pesticide, during inoculation cultured in vitro, do not need directly to contact flower pesticide and culture matrix, avoid contact stain, reduce pollution probability yet.
(3) the present invention is in flower pesticide collection process, by regulating frequency of oscillation, controls flower pesticide falling speed, the collection of flower pesticide is more flexible, without the need to tweezers picking flower pesticide from clever shell, contribute to relieving eye strain and operating personnel's continuous firing state, operate easier.
Accompanying drawing explanation
Fig. 1 is the schematic diagram that embodiment 1 ~ 2 receiving flask collects flower pesticide.
Fig. 2 is the callus that embodiment 2 anther culture is formed.
Embodiment
Below in conjunction with specific embodiment, the present invention is further explained.
Fig. 1 shows the schematic diagram that embodiment 1 ~ 2 receiving flask collects flower pesticide, visible in figure, and receiving flask is cylindric, and be divided into top cylinder, doffing two-layer, top cylinder and doffing are by being threaded.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 clamping the stem stalk of spike of rice 6, the bottom of doffing 3 is loaded with agar filter paper 4.When collecting flower pesticide, whole receiving flask is placed on oscillator 5, and flower pesticide shakes by the vibration of oscillator 5.
Embodiment 1
In receiving flask, the method for Rice Anther is collected in vibration, comprises the following steps:
(1) the rice bud sword-like leave pulvinus of extracting out is to the 2nd leaf, and spike of rice grain husk shell gets fringe in field when there is light green color, by spike of rice in 4 degrees Celsius of Cold pretreatment 5 days.
(2) spike of rice after process is placed in sterile tissue blake bottle, the alcohol with 70% carries out surface sterilization 1min, removes alcohol, with the hypochlorite disinfectant 15min of 1% in single aseptic superclean bench, removes clorox, with aseptic water washing 5 times.
(3) spike of rice after sterilization is cut off 1/3 of clever shell top with upper part with scissors in single aseptic super-clean bench, in Fig. 1, show complete clever shell 6, remove the clever shell 7 on top.
(4) aseptically receiving flask bottom is set off sterile letheen filter paper, to cut off the fringe portion of clever shell down, stem stalk puts into receiving flask upward, tightens upper and lower two layers of receiving flask (i.e. top cylinder, doffing), and stem stalk is clipped, the distance between at the bottom of the top in fringe portion and receiving flask bottle is 3cm.
(5) receiving flask puts into vibration collection on oscillator, and frequency of oscillation regulates at 3 hertz, and vibration 15s, flower pesticide is collected in receiving flask substantially.
(6) in aseptic super-clean bench, turn on receiving flask after having collected, take out the agar filter paper of bottle bottom, the agar filter paper with flower pesticide being transferred in culture dish, being placed in 25 DEG C of incubator light culture to inducing callus.
24 bottles of flower pesticide within two hours, can be collected, and every bottle of flower pesticide >=150 piece, cultivated by the flower pesticide of collection, light culture is after 35 days, and statistics callus induction rate, its inductivity is 10.16%, and does not produce any pollution.
Embodiment 2
In receiving flask, the method for Rice Anther is collected in vibration, comprises the following steps:
(1) when the rice bud sword-like leave pulvinus of extracting out is to the 2nd leaf, spike of rice grain husk shell gets fringe in field when there is light green color, by spike of rice in 5 degrees Celsius of Cold pretreatment 6 days.
(2) spike of rice after process is placed in sterile tissue blake bottle, the alcohol with 70% carries out surface sterilization 1min, removes alcohol, with the hypochlorite disinfectant 15min of 1% in single aseptic superclean bench, removes clorox, with aseptic water washing 5 times.
(3) cut off after the spike of rice sterilization after process 1/2 of clever shell top with upper part with scissors in the clean platform of double aseptic behaviour.
(4) in the clean platform of aseptic behaviour, receiving flask bottom is set off aseptic agar filter paper, to cut off the fringe portion of clever shell down, stem stalk puts into receiving flask upward, and is clipped by stem stalk, tighten upper and lower two layers of receiving flask (i.e. top cylinder, doffing), the distance between at the bottom of the top of spike of rice and receiving flask bottle is 5cm.
(5) receiving flask puts into vibration collection on oscillator, and frequency of oscillation regulates at 10 hertz, and duration of oscillation is 8s.
(6) in the clean platform of aseptic behaviour, turn on receiving flask after having collected, take out the agar filter paper of bottle bottom, flower pesticide being transferred in culture dish, being placed in 25 DEG C of incubator light culture to inducing callus.
26 bottles of flower pesticide within two hours, can be collected, and every bottle of flower pesticide >=150 piece, light culture is after 25 days, and statistics callus induction rate, its inductivity is 10.35%, and does not produce any pollution, and Fig. 2 shows the callus that the present embodiment anther culture is formed.
Comparative example
(1) the rice bud sword-like leave pulvinus of extracting out is to the 2nd leaf, and spike of rice grain husk shell gets spike of rice in field when there is light green color, by spike of rice in 5 degrees Celsius of Cold pretreatment 6 days.
(2) spike of rice after process is placed in sterile tissue blake bottle, the alcohol with 70% carries out surface sterilization 1min, removes alcohol, with the hypochlorite disinfectant 15min of 1% in single aseptic superclean bench, removes clorox, with aseptic water washing 5 times.
(3) conventional method is adopted when getting flower pesticide, first ripped by the clever shell of eye scissors by spike of rice, take out flower pesticide and be placed in sterile petri dish, have after some until flower pesticide, being inoculated in medium with ophthalmic tweezers, being placed in 25 DEG C of incubator light culture to inducing callus.
Through statistics, people's operation gets 1200 pieces of flower pesticide, about 2 hours, is inoculated in medium by these 1200 pieces of flower pesticide, each culture dish inoculates 150 pieces, totally 8 culture dishes, cultivates after 35 days, statistics callus induction rate, its inductivity is 9.95%, and finds that there is two wares generation pollutions.
Claims (5)
1. a collection method for Rice Anther, comprising:
Get spike of rice, after sterilization, remove the top of clever shell; By rice bud stem stalk upward, fringe portion makes described fringe portion be suspended in receiving flask down, and vibration is collected and obtained described flower pesticide;
Described receiving flask is divided into two-layer up and down, and upper strata is provided with the fixture for fixing described spike of rice, and lower floor is loaded with agar filter paper; Distance between at the bottom of described fringe portion and receiving flask bottle is 2 ~ 6cm;
The frequency of described vibration is 0.1 ~ 10 hertz; The time of described vibration is 1 ~ 30s.
2. collection method as claimed in claim 1, is characterized in that, the sword-like leave pulvinus that rice bud is extracted out, to the 2nd leaf, takes out spike of rice when light green color appears in clever shell from described rice bud.
3. collection method as claimed in claim 1, is characterized in that, before sterilization, is placed 4 ~ 6 days by described spike of rice in 4 ~ 6 DEG C.
4. collection method as claimed in claim 1, it is characterized in that, the method for described sterilization comprises: by described spike of rice with after 70% alcohol disinfecting 30s ~ 1min, and clorox spike of rice being placed in 1% 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.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| KR20080050759A (en) * | 2006-12-04 | 2008-06-10 | 임장혁 | Flower pine pick catch system |
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
|---|
| 水稻花药愈伤组织诱导的多因子正交试验研究;王辉等;《广西植物》;20110731;第31卷(第4期);第545-549页 * |
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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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