CN112314434A - 一种彩色矮丛盆栽水稻的分子育种方法 - Google Patents

一种彩色矮丛盆栽水稻的分子育种方法 Download PDF

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CN112314434A
CN112314434A CN202011513827.4A CN202011513827A CN112314434A CN 112314434 A CN112314434 A CN 112314434A CN 202011513827 A CN202011513827 A CN 202011513827A CN 112314434 A CN112314434 A CN 112314434A
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沈雨民
熊文涛
陈明亮
熊焕金
吴小燕
罗世友
肖叶青
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RICE RESEARCH INSTITUTE OF JIANGXI ACADEMY OF AGRICULTURAL SCIENCES
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Abstract

本发明公开了一种彩色矮丛盆栽水稻的分子育种方法：通过选用籼型矮杆多分蘖突变体作为轮回亲本，与紫叶稻突变体、黄绿叶突变体分别杂交获得F1；F1代继续分别与轮回亲本杂交，获BC1F1；BC1F1代苗期提取叶片DNA，利用连锁分子标记分别对矮杆多分蘖基因、紫叶基因和黄绿叶基因进行检测，挑选含目标基因的单株继续与轮回亲本杂交获得BC2F1；继续经过多次分子标记选择回交，直至获得BC6F1；自交繁代，在后代中挑选紫叶矮杆多分蘖、黄绿叶矮杆多分蘖株系，与轮回亲本组成彩色水稻。由于轮回亲本是籼型矮杆多分蘖突变体，通过本方法培育出的株系，具有小巧玲珑、分蘖多、株型稍披散、颜色多样等特点，特别适合用于室内盆栽。

Description

一种彩色矮丛盆栽水稻的分子育种方法
技术领域
本发明属于景观水稻育种和分子遗传学技术领域，具体涉及一种彩色矮丛盆栽水稻的分子育种方法。
技术背景
中国是农业古国，历史悠久的农耕文化让我们引以为豪。水稻作为中华民族的主要农作物之一，养育了千千万万的中华人民。水稻种植历史长远，距今七千年前的浙江河姆渡遗址就出现了以水稻种植为主的农业部落。伴随着农村城镇化的发展及农村人口向城镇的迁移，部分人特别是年轻人可能已经无法辨认水稻，更遑论水稻的生长过程。盆栽作为城市室内主要的绿植，具有净化室内空气和陶冶人们情操等作用。如果将水稻盆栽种植到室内，不仅可以普及水稻知识，而且可以加深城市居民对稻作文化的了解。
然而，普通水稻在室内由于相对高大，色彩单一，缺乏观赏性，不适宜在室内种植。
发明内容
为解决上述问题，本发明的目的在于提供一种彩色矮丛盆栽水稻的分子育种方法。通过本发明选用籼型矮杆多分蘖突变体作为轮回亲本，与紫叶稻突变体、黄绿叶突变体分别杂交、多轮回交，利用分子标记快速选择，选育出色彩鲜明且便于在室内种植的盆栽观赏水稻。
本发明的技术方案如下：
一种彩色矮丛盆栽水稻的分子育种方法，其特征在于，包括以下步骤：
(1)以籼型矮杆多分蘖突变体作为轮回亲本，以紫叶突变体和黄绿叶突变体作为供体亲本，分别杂交获得杂种F1；
(2)种植F1，继续与轮回亲本回交获得BC1F1；
(3)种植BC1F1，苗期提取叶片DNA，利用连锁分子标记分别对矮杆多分蘖基因、紫叶基因和黄绿叶基因进行检测，挑选含目标基因的单株继续与轮回亲本杂交获得BC2F1；
(4)继续步骤(3)获得BC3F1，BC4F1，BC5F1，BC6F1；
(5)种植BC6F1，自交繁代，在后代中挑选紫叶矮杆多分蘖、黄绿叶矮杆多分蘖株系，与轮回亲本组成彩色矮丛盆栽水稻。
优选的，步骤(1)所述的籼型矮杆多分蘖突变体为九香占的D10基因突变体；紫叶突变体为一未克隆突变体PLS；黄绿叶突变体为Ygl8基因突变体。
优选的，步骤(3)所述的连锁分子标记，包括D10基因连锁SSR分子标记RM1095，PLS基因连锁SSR分子标记RM1302,Ygl8基因连锁SSR分子标记RM6321。
优选的，所述的RM1095
其左端序列为CCCATTCAGTTGATCCTGTCTGC
右端序列为AGCTGGGATGCAGAAGAGTATGG
所述的RM1302
其左端序列为GAACGTGAAAGAGAAGCTCATGG
右端序列为TCTCCTTCTCTCCCAACATCTCG
所述的RM6321
其左端序列为CCCAGGAATATCTCTATACACACAGC
右端序列为CTCTACCTCGCTGTTGTCTCTGC。
有益效果：
(1) 在彩色矮丛盆栽水稻的培育过程中，利用目标性状的连锁分子标记在苗期进行分子标记辅助选择，加快了选育进度。
(2) 所培育的彩色矮丛盆栽水稻，具有不仅能丰富盆栽种类，而且具有趣味性和普及水稻知识的功能。克服了相对高大的水稻在室内不便种植的缺点。
附图说明
图1为彩色矮丛盆栽水稻的培育流程图；
图2为D10基因连锁分子标记RM1095在BC1F1代丙烯酰胺凝胶的检测胶图；M为Marker，1为D10突变体，2为Ygl8突变体，3-24为D10突变体/Ygl8突变体的BC1F1群体单株（其中7,11,12,14,18号单株在D10基因位点为杂合型，其余单株携带纯合D10突变基因）；
图3为PLS基因连锁分子标记RM1302在BC1F1代丙烯酰胺凝胶的检测胶图；M为Marker，1为D10突变体，2为PLS突变体，3-24为D10突变体/PLS突变体的BC1F1群体单株（其中4,9,19,21,22,23,24号单株为杂合型，携带PLS突变基因；其余单株不含PLS突变基因）；
图4为Ygl8基因紧密连锁标记RM6321在BC1F1代丙烯酰胺凝胶的检测胶图；M为Marker，1为D10突变体，2为Ygl8突变体，3-24为D10突变体/Ygl8突变体的BC1F1群体单株（其中14,16,19,20号单株为杂合型，携带Ygl8突变基因；其余单株不含Ygl8突变基因）；
图5为最终选育成的彩色矮丛盆栽水稻，株高约为55cm，分蘖数20左右；从左至右分别为矮杆多分蘖黄绿叶株系，矮杆多分蘖紫叶株系，矮杆多分蘖株系。
具体实施方式
下面结合附图和具体实施例对本发明作进一步说明，但本发明并不限于以下实施例。所述方法无特别说明均为常规方法，所述原材料如无特别说明均能从公开商业途径获得。
（1）以籼稻品种九香占（D10基因突变）作为轮回亲本，以PLS突变体和Ygl8突变体作为供体亲本，分别杂交获得F1；
（2）种植F1，继续与轮回亲本回交获得BC1F1；
（3）种植BC1F1，苗期提取叶片DNA，利用D10基因连锁SSR分子标记RM1095(其左端序列为CCCATTCAGTTGATCCTGTCTGC，右端序列为AGCTGGGATGCAGAAGAGTATGG)，PLS基因连锁SSR分子标记RM1302(其左端序列为GAACGTGAAAGAGAAGCTCATGG，右端序列为TCTCCTTCTCTCCCAACATCTCG),Ygl8基因连锁SSR分子标记RM6321(其左端序列为CCCAGGAATATCTCTATACACACAGC，右端序列CTCTACCTCGCTGTTGTCTCTGC)分别对D10基因、PLS基因和Ygl8基因进行检测（图2,3,4），挑选含目标基因的单株继续与轮回亲本杂交获得BC2F1；
(4) 继续步骤(3)获得BC3F1，BC4F1，BC5F1, BC6F1;
(5) 种植BC6F1, 自交繁代，在后代中挑选紫叶矮杆多分蘖、黄绿叶矮杆多分蘖株系，与轮回亲本组成彩色盆栽水稻（图5）。
以上对本发明的具体实施例进行了详细描述，但本发明的保护范围并不局限于此，任何熟悉本技术领域的技术人员在本发明揭露的技术范围内，根据本发明的技术方案及其发明构思加以等同替换或改变，都应涵盖在本发明的保护范围之内。
序列表
<110> 江西省农业科学院水稻研究所
<120> 一种彩色矮丛盆栽水稻的分子育种方法
<141> 2020-12-18
<160> 6
<170> SIPOSequenceListing 1.0
<210> 1
<211> 23
<212> DNA
<213> D10基因连锁SSR分子标记RM1095(左端)
<400> 1
cccattcagt tgatcctgtc tgc 23
<210> 2
<211> 23
<212> DNA
<213> D10基因连锁SSR分子标记RM1095(右端)
<400> 2
agctgggatg cagaagagta tgg 23
<210> 3
<211> 23
<212> DNA
<213> PLS基因连锁SSR分子标记RM1302(左端)
<400> 3
gaacgtgaaa gagaagctca tgg 23
<210> 4
<211> 23
<212> DNA
<213> PLS基因连锁SSR分子标记RM1302(右端)
<400> 4
tctccttctc tcccaacatc tcg 23
<210> 5
<211> 26
<212> DNA
<213> Ygl8基因连锁SSR分子标记RM6321(左端)
<400> 5
cccaggaata tctctataca cacagc 26
<210> 6
<211> 23
<212> DNA
<213> Ygl8基因连锁SSR分子标记RM6321(右端)
<400> 6
ctctacctcg ctgttgtctc tgc 23

Claims (6)

1.一种彩色矮丛盆栽水稻的分子育种方法，其特征在于，所述方法包括以下步骤：
(1)以籼型矮杆多分蘖突变体作为轮回亲本，以紫叶突变体和黄绿叶突变体作为供体亲本，分别杂交获得杂种F1；
(2)种植F1，继续与轮回亲本回交获得BC1F1；
(3)种植BC1F1，苗期提取叶片DNA，利用连锁分子标记分别对矮杆多分蘖基因、紫叶基因和黄绿叶基因进行检测，挑选含目标基因的单株继续与轮回亲本杂交获得BC2F1；
(4)继续步骤(3)获得BC3F1，BC4F1，BC5F1，BC6F1；
(5)种植BC6F1，自交繁代，在后代中挑选紫叶矮杆多分蘖、黄绿叶矮杆多分蘖株系，与轮回亲本组成彩色矮丛盆栽水稻。
2.根据权利要求1所述的一种彩色矮丛盆栽水稻的分子育种方法，其特征在于，步骤(1)所述的籼型矮杆多分蘖突变体为九香占的D10基因突变体；紫叶突变体为一未克隆突变体PLS；黄绿叶突变体为Ygl8基因突变体。
3.根据权利要求1所述的一种彩色矮丛盆栽水稻的分子育种方法，其特征在于步骤(3)所述的连锁分子标记，包括D10基因连锁SSR分子标记RM1095，PLS基因连锁SSR分子标记RM1302，Ygl8基因连锁SSR分子标记RM6321。
4.根据权利要求3所述的一种彩色矮丛盆栽水稻的分子育种方法，其特征在于：所述的D10基因连锁SSR分子标记RM1095
其左端序列为CCCATTCAGTTGATCCTGTCTGC
右端序列为AGCTGGGATGCAGAAGAGTATGG
所述的PLS基因连锁SSR分子标记RM1302
其左端序列为GAACGTGAAAGAGAAGCTCATGG
右端序列为TCTCCTTCTCTCCCAACATCTCG
所述的Ygl8基因连锁SSR分子标记RM6321
其左端序列为CCCAGGAATATCTCTATACACACAGC
右端序列为CTCTACCTCGCTGTTGTCTCTGC。
5.根据权利要求1所述的一种彩色矮丛盆栽水稻的分子育种方法，其特征在于：步骤（5）所述的紫叶矮杆多分蘖、黄绿叶矮杆多分蘖株系，其分蘖数达到20个左右，株高55cm左右。
6.根据权利要求1-5任一所述的一种彩色矮丛盆栽水稻的分子育种方法，其特征在于：用于彩色矮丛盆栽水稻育种。
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Cited By (1)

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CN114831017A (zh) * 2021-12-28 2022-08-02 浙江大学 一种家庭阳台盆栽用水稻的选育方法

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