CN101176424B - 提高栽培稻×非aa型野生稻成胚率的新方法 - Google Patents
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Abstract
本发明属于植物杂交育种技术。为解决栽培稻同非AA型野生稻种间远缘杂交不亲和的难题并提高成胚率特提出本方案。首先通过抽穗开花期温湿度调节和应用挡风设施，获得大量高活力的非AA型野生稻新鲜花粉，然后利用深色套袋或温汤处理调节父母本花时相遇，用镊子夹取单个小花的花药或将小穗置于待授粉母本稻穗上用手抖动，可提高非AA型野生稻的花粉利用率，重复授粉2～3次效果更好。利用各种类型的栽培稻材料与非AA型野生稻杂交，筛选亲和性好的材料与非AA型野生稻杂交。此方法的操作简单，成本低，能使成胚率由0～40％稳定提高到30％～60％。此法较好的解决了栽培稻同非AA型野生稻种间远缘杂交不亲和的难题，显著提高了成胚率。
Description
技术领域：本发明专利属于植物杂交育种技术。它具体涉及一种能显著提高栽培稻×非AA型野生稻的杂交成胚率的新方法。
背景技术：非AA型野生稻涵盖B、C、D、E、F、G、H、J、K九大基因组，广泛分布于世界各地，具有丰富的遗传多样性和许多优良性状，是一座天然的水稻基因宝库，可以为拓宽水稻遗传基础提供丰富的种质资源。如染色体组型为CCDD的重颖野生稻(四倍体)，具有抗稻瘟病、抗菲律宾水蝇、螟虫和褐飞虱等病虫害的特性，有高耐寒、耐荫蔽、弱光条件下生长较好的特点，在水稻育种上有重大利用价值。但是，迄今为止，由于非AA型野生稻与栽培稻的亲缘关系较远，遗传差异很大，存在一定程度的生殖隔离，野栽交表现不亲和，加上非AA型野生稻的开花习性不同于栽培稻和普通野生稻，如开花不整齐，一天一个穗子只开几朵、花时不遇等，技术人员到田间做杂交时难以获得足够的非AA型野生稻新鲜花粉，致使它与栽培稻或普通野生稻杂交成胚率低、难度大、效率低。
发明内容：本发明的目的是为了解决栽培稻同非AA型野生稻种间远缘杂交不亲和的难题，提高栽培稻同非AA型野生稻种间远缘杂交的成功率，特提出本方案——提高栽培稻×非AA型野生稻成胚率的新方法，它用调整非AA型野生稻播期、遮光等方法使杂交父本母本的花期相遇，套袋、温汤处理使花时相遇，提高成胚率并降低技术难度和研究成本。
按如上构思，本方案提出的提高栽培稻×非AA型野生稻成胚率的新 方法，其特征是：
①将非AA型野生稻经播期安排和遮光处理调控到25～32℃温度条件下抽穗开花，以获得大量高活力的野生稻新鲜花粉，提高受精机率；
②建立塑料棚简易挡风设施防止花粉散失，提高获得野生稻新鲜花粉的数量，主 要针对田间条件下花粉易散失难以获得大量花粉的非AA型野生稻种，如重颖野生稻；
③给即将开花的非AA型野生稻小穗套深色牛皮纸袋延迟开花散粉，或用温水43℃～45℃处理稻穗3～5分钟使栽培稻亲本提前开花，使双亲花时相遇，提高杂交成功率；
④非AA型野生稻开花时，用镊子夹取单个小花的花药或用剪刀剪取小穗置于待授粉母本稻穗上，用手抖动小花或小穗授粉，提高非AA型野生稻的花粉利用率；
⑤通过杂交筛选出与非AA型野生稻亲和性好的栽培稻材料，再利用亲和性好的材料与非AA型野生稻杂交，提高杂交的成功率。
在本方案中，将栽培稻各类型中具有代表性的材料与非AA型野生稻进行广泛杂交，统计成胚率，选取成胚率高的类型与非AA型野生稻杂交。采用本方案能体现如下的优越性：一是不需要昂贵的仪器和设备，成本低；二是方法简单，技术难度小，容易掌握；三是广泛适宜于利用野生稻进行远缘杂交的水稻遗传育种种质资源创新研究。只需要将育种工作者平时用到的技术和设备巧妙调整、组装，就能大幅度提高栽培稻×非AA型野生稻种间远缘杂交的成胚率，为充分挖掘和利用非AA型野生稻打下基础。
具体实施方式：首先，必须了解非AA型野生稻的形态生理特性，包括开花习性(花时、粉量、散粉特性)、高低温耐性、感光性等。采用遮光处理、分期播种等措施将非AA型野生稻和栽培稻亲本的花期调至最适宜非AA型野生稻开花散粉、栽培稻开花受精的温度条件下(25℃-32℃)， 也可借助人工气候室等控温设备进行温度调节，从而不受季节限制，均可获得大量高活力的野生稻新鲜花粉。如野生稻和栽培稻花时不遇，利用深色牛皮纸袋套住将开花的小穗可延迟开花，开花后花粉也不易散失；利用温水处理(37℃)可提前开花，且开花整齐。非AA型野生稻花期田间自然风力较大的地方，需要搭建塑料棚等简易挡风设施，防止开花后花粉很快就被吹走。野生稻一般每个花药的花粉量大，但开花不整齐，一个穗子一天只开几朵，即使剪颖也难以改变其固有特性，可利用镊子夹取单个小花的花药授粉，或用剪刀剪取小穗置于待授粉稻穗上，用手抖动小穗授粉，可大幅度提高野生稻的花粉利用率，减少野生稻的种植量。授粉前1天将用作母本的栽培稻和普通野生稻剪颖去雄后套袋，在非AA型野生稻开花时直接授粉，或采取措施使非AA型野生稻与栽培稻亲本花期相遇，待母本小穗开花较多时授粉，一般重复授粉2-3次。通过广泛杂交筛选出亲和性较好(具体操作时可将重点放在广亲和材料上)的栽培稻亲本，利用亲和性好的材料与非AA型野生稻杂交可将大量新鲜花粉授到栽培稻母本高活力的柱头上，大幅度提高了栽培稻×非AA型野生稻种间远缘杂交的成胚率，解决野栽交表现不亲和的难题。
下面分别叙述实施例的具体情况：
在海南进行栽培稻×非AA型重颖野生稻的种间远缘杂交，将杂交安排在3月中旬以前，3月下旬以后，一般温度较高，重颖野生稻将散粉不畅，难以得到足够的花粉。海南风大，重颖野生稻开花后花粉很快就被吹走，因此需要搭建塑料棚等简易挡风设施。为了减少挡风设施对野生稻的生长产生影响，以简易塑料棚为例，首先，塑料棚最好能搭建得较大，以 不与野生稻茎秆叶片接触为宜。其次，塑料膜不能固定死，白天能撤除，只留竹竿，晚上重新遮盖起来。用深色牛皮纸袋套住穗子花粉也不易散失，且可延迟授粉时间，但由于套袋后野生稻茎秆易被风吹断，需用竹竿等将已套袋的穗子固定。在授粉前一天下午将用作母本的栽培稻剪颖去雄，在第二天上午重颖野生稻开花散粉时，用镊子夹取重颖野生稻小穗上的花药或用剪刀剪取开花的小穗置于待授粉稻穗上，用手抖动小穗即可。如果技术熟练，重颖野生稻每个颖花的花粉可以授1～2个稻穗，即使需要做较多杂交，有2～3棵长势较好的野生稻就可提供足够的花粉，一般情况下，有1棵长势较好的野生稻就足够了。重复授粉2～3次效果更好。如果再选用 亲和性较好的栽培稻作亲本，如F4911，结实率可达到40％～60％。
在长沙进行栽培稻×重颖野生稻的种间远缘杂交，重颖野生稻感光性强，在长沙需要做短光处理，自然条件下野生稻最佳抽穗散粉期为9月中、下旬，借助温室可将杂交安排在9月下旬到10月上旬，其它方面与前者 相同。
通过上述方法操作，较好的解决了栽培稻同非AA型野生稻种间远缘杂交不亲和的难题，显著提高了栽培稻同非AA型野生稻种间远缘杂交的成胚率，由0～40％稳定提高到30％～60％。为充分挖掘和利用远缘物种进行水稻遗传育种种质资源创新提供了新方法，为其它作物遗传育种种质资源创新提供了的借鉴经验。
Claims (2)
1.一种提高栽培稻×非AA型野生稻成胚率的新方法，其特征是：
①通过播期安排和遮光处理措施将非AA型野生稻调控到适宜温度条件下，一般为25℃～32℃抽穗开花，以获得大量高活力的野生稻新鲜花粉，提高受精机率；
②建立塑料棚简易挡风设施防止花粉散失，提高获得野生稻新鲜花粉的数量；
③给即将开花的非AA型野生稻小穗套深色牛皮纸袋延迟开花散粉，或用温水43℃～45℃处理稻穗3～5分钟使栽培稻亲本提前开花，使双亲花时相遇，提高杂交成功率；
④非AA型野生稻开花时，用镊子夹取单个小花的花药或用剪刀剪取单个小穗置于待授粉母本稻穗上，用手抖动小花或小穗授粉，提高非AA型野生稻的花粉利用率；
⑤通过广泛杂交筛选出与非AA型野生稻亲和性好的栽培稻材料，再利用亲和性好的材料与重颖野生稻杂交，提高杂交的成功率。
2.根据权利1所要求所述的新方法，其特征是：将栽培稻各类型中具有代表性的材料与非AA型野生稻进行广泛杂交，统计成胚率，选取成胚率高的类型与非AA型野生稻杂交。
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| 吴妙燊等.野生稻种质利用技术的研究——Ⅰ.稻属种间杂种(AA×CC染色体组)的获得.《中国种业》.1988,(第01期),6-9. * |
| 张武汉,等.非AA 型野生稻资源在水稻育种中的利用.《杂交水稻》.2006,第21卷(第5期),1-7. * |
| 杨长桃等.延续授粉克服水稻远缘杂交不实的探讨.《福建农业科技》.1989,(第03期),7-8. * |
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