甘蓝型油菜抗（耐）菌核病PolCMS恢复系的选育
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摘要
我国是世界上最大的油菜生产国,2000年以来种植面积已突破7,337,000公顷,面积和总产均占世界的三分之一,长江流域是世界油菜最大的产区,面积约占世界四分之一。油菜既是高油分作物,又是高蛋白作物,在世界范围如加拿大、西欧、北欧、澳大利亚、印度等都有着广泛的种植面积,是世界上食用植物油和植物蛋白的主要来源之一。油菜菌核病是制约我国油菜生长的重要病害,影响产量很大。尤其是长江中下游油菜主产区,生育后期遭遇高温多雨天气,菌核病发生严重。因此,选育抗菌核病油菜品种及资源是当务之急。
本研究以小孢子培养和分子标记辅助选择两种途径来选育抗病的恢复系。一方面,以抗菌核病的恢复系中双4号R DH和不抗病的恢复系5148 DH、不育系109A为基础材料,利用小孢子培养,获得抗感分离的DH植株,通过田间抗病鉴定,得到抗病的新恢复系。另一方面,我们利用分子标记辅助选择,对优良材料的抗性进行改良,选择遗传背景与轮回亲本一致同时在抗病鉴定中又表现抗病的植株,获得抗病的新恢复系。主要研究结果如下:
1.通过对(5148 DH×中双4号R DH)的F_1和(109 A×中双4号R DH)的F_1代做小孢子培养,分别获得438和172个DH株系。
2.在对两个DH群体做苗期抗病鉴定时,DH-1和DH-2两个群体分别获得69个和39个抗病的株系。
3.对两个DH群体做成株期抗病鉴定,分别获得38个和17个抗病个株系;同时发现3d和5d、5d和7d、3d和7d的病斑长度的相关性较高,在两个群体中均未发现抗侵入和抗扩展的株系。
4.在两个DH群体中共获得9株苗期和成株期均表现抗性的单株,苗期和成株期的抗病性相关性不高。
5.利用分子标记辅助选择,通过两次回交和背景选择,获得遗传背景与5148DH一致,同时又表现抗病性的单株4株,可以作为新的恢复系种质资源。
China is the largest nation for Brassica napus production in the world. The area of the planting has exceeded 7,337,000 hm~2 and both the area and the total amount are one third of the entire production of the world. The drainage area of the Yangtze River provides the largest yield area of Brassica napus all over the world, which almost occupies one fourth of the whole earth. Brassica napus contains both high oil and high protein and it is widely cultivated in Canada, West Europe, North Europe, Australia, India and so on. Brassica napus is also one of the main resource of edible oil and plant protein in the world.
Sclerotinia sclerotiorum, which is one kind of serious diseases that confines development of Brassica napus and causes significant production reduction. Particularly, it is during the period of high temperature and rainy days that the Brassica napus come on disease more seriously on its later growth phase in center down main yield areas of Yangtze River. Thus, it is exigent and desirable to select disease-resistant new varieties and germplasm resources.
This study focus on selection of disease-resistant restorer lines through two approaches of microspore culture and marker assisted selection. On the one hand, we derives disease-resistant novel restorers on the basis of disease-resistant restorer ZHONG SHUANG 4 RDH and non-resistant restorer 5148 DHL sterility 109A, using microspore culture of Brassica napus to obtain doubled haploid (DH) lines of separation population between resistant and sensitive disease plants and through identification of disease-resistant in the field. On the other hand, in order to derive novel disease-resistant restorer lines, we use marker assisted selection to improve resistance property of smart materials and select materials both having genetic background consistent with recurrent parent and display disease-resistance in identification of disease-resistant experiment. The results are as follows:
1. Through microspore culture for F_1 generation of (5148 DH×ZHONGSHUANG 4 RDH) and (109A×ZHONGSHUANG 4 RDH), we derived 438 and 171 different genotype DH single plants respectively.
2. When identification of disease-resistant for 2 DH populations in the seedling stage, we obtained 69 and 39 different genotype disease-resistant single plants respectively.
3. When identification of disease-resistant for 2 DH populations in the adult plant stage, we derived respectively 38 and 17 different genotype disease-resistant single plants; Simultaneously there were quite correlation on length of disease spot between 3d and5d, 5d and 7d,3d and 7d. but we could not find any single plant of immunity and resistant enlargement for Sclerotinia sclerotiorum.
4. There were 9 single plants, which showed resistance property both on seedling stage and adult plant stage. And there was no significant correlation on disease-resistance between seedling stage and adult plant stage.
5. There were 4 single plants, which both had the same genetic background with 5148 DH using marker assisted selection and through backcross twice and selfing once and displayed disease-resistance. These single plants might be as novel restorer lines germplasm resources.
本研究以小孢子培养和分子标记辅助选择两种途径来选育抗病的恢复系。一方面,以抗菌核病的恢复系中双4号R DH和不抗病的恢复系5148 DH、不育系109A为基础材料,利用小孢子培养,获得抗感分离的DH植株,通过田间抗病鉴定,得到抗病的新恢复系。另一方面,我们利用分子标记辅助选择,对优良材料的抗性进行改良,选择遗传背景与轮回亲本一致同时在抗病鉴定中又表现抗病的植株,获得抗病的新恢复系。主要研究结果如下:
1.通过对(5148 DH×中双4号R DH)的F_1和(109 A×中双4号R DH)的F_1代做小孢子培养,分别获得438和172个DH株系。
2.在对两个DH群体做苗期抗病鉴定时,DH-1和DH-2两个群体分别获得69个和39个抗病的株系。
3.对两个DH群体做成株期抗病鉴定,分别获得38个和17个抗病个株系;同时发现3d和5d、5d和7d、3d和7d的病斑长度的相关性较高,在两个群体中均未发现抗侵入和抗扩展的株系。
4.在两个DH群体中共获得9株苗期和成株期均表现抗性的单株,苗期和成株期的抗病性相关性不高。
5.利用分子标记辅助选择,通过两次回交和背景选择,获得遗传背景与5148DH一致,同时又表现抗病性的单株4株,可以作为新的恢复系种质资源。
China is the largest nation for Brassica napus production in the world. The area of the planting has exceeded 7,337,000 hm~2 and both the area and the total amount are one third of the entire production of the world. The drainage area of the Yangtze River provides the largest yield area of Brassica napus all over the world, which almost occupies one fourth of the whole earth. Brassica napus contains both high oil and high protein and it is widely cultivated in Canada, West Europe, North Europe, Australia, India and so on. Brassica napus is also one of the main resource of edible oil and plant protein in the world.
Sclerotinia sclerotiorum, which is one kind of serious diseases that confines development of Brassica napus and causes significant production reduction. Particularly, it is during the period of high temperature and rainy days that the Brassica napus come on disease more seriously on its later growth phase in center down main yield areas of Yangtze River. Thus, it is exigent and desirable to select disease-resistant new varieties and germplasm resources.
This study focus on selection of disease-resistant restorer lines through two approaches of microspore culture and marker assisted selection. On the one hand, we derives disease-resistant novel restorers on the basis of disease-resistant restorer ZHONG SHUANG 4 RDH and non-resistant restorer 5148 DHL sterility 109A, using microspore culture of Brassica napus to obtain doubled haploid (DH) lines of separation population between resistant and sensitive disease plants and through identification of disease-resistant in the field. On the other hand, in order to derive novel disease-resistant restorer lines, we use marker assisted selection to improve resistance property of smart materials and select materials both having genetic background consistent with recurrent parent and display disease-resistance in identification of disease-resistant experiment. The results are as follows:
1. Through microspore culture for F_1 generation of (5148 DH×ZHONGSHUANG 4 RDH) and (109A×ZHONGSHUANG 4 RDH), we derived 438 and 171 different genotype DH single plants respectively.
2. When identification of disease-resistant for 2 DH populations in the seedling stage, we obtained 69 and 39 different genotype disease-resistant single plants respectively.
3. When identification of disease-resistant for 2 DH populations in the adult plant stage, we derived respectively 38 and 17 different genotype disease-resistant single plants; Simultaneously there were quite correlation on length of disease spot between 3d and5d, 5d and 7d,3d and 7d. but we could not find any single plant of immunity and resistant enlargement for Sclerotinia sclerotiorum.
4. There were 9 single plants, which showed resistance property both on seedling stage and adult plant stage. And there was no significant correlation on disease-resistance between seedling stage and adult plant stage.
5. There were 4 single plants, which both had the same genetic background with 5148 DH using marker assisted selection and through backcross twice and selfing once and displayed disease-resistance. These single plants might be as novel restorer lines germplasm resources.
引文
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