大豆叶茸毛着生状态与筛豆龟蝽抗性的关联及基因定位
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摘要
作物的抗虫性常常与作物的形态特征有关,茸毛着生状态这一形态特征是昆虫寻觅适宜生栖环境和食物的感觉依据,影响着昆虫在植物表面的取食和产卵等行为。已有研究结果指出叶片茸毛的着生状态与大豆抗虫性存在显著相关,发现叶片上具匍匐状茸毛的大豆材料对豆卷叶螟的抗性好于叶片直立茸毛的大豆。经典遗传学研究表明大豆叶片茸毛着生状态受少数基因控制,但该特性也表现较典型的数量性状特点,受基因型、生长时期及环境条件等因素的影响较大。筛豆龟蝽通过刺吸大豆茎叶以及引发病害而对大豆的产量和品质造成巨大危害,近年来已成为我国南方地区的主要害虫,且有上升的趋势。选育抗虫品种是防治筛豆龟蝽的最重要措施,揭示大豆资源中抗性遗传变异、发掘抗虫基因资源是抗虫育种的首要工作。
本研究利用来源于全国24个省份的400份大豆地方品种研究大豆叶茸毛着生状态与筛豆龟蝽抗性的关系,比较叶片和叶柄茸毛着生状态与筛豆龟蝽抗性的关联；进一步通过重组自交系群体和构建的高代自交分离群体,对控制叶片茸毛着生状态的基因进行定位和验证,为叶片茸毛着生状态基因的育种利用及其与抗虫性的相互作用等研究提供指导。本研究的主要结果如下：
(1)大豆种质资源品种间对筛豆龟蝽抗性存在较大的变异,筛豆龟蝽的抗性等级与生态区间也存在显著的相关,东北、黄淮海生态区的高感品种的频率高于长江中下游,中南,西南高原和华南热带等南方生态区,而长江中下游,中南,西南高原和华南热带等南方生态区高抗品种的频率高于东北、黄淮海生态区。
(2)叶茸毛着生状态与筛豆龟蝽的危害率两者间存在极显著关联。匍匐型叶片茸毛抗筛豆龟蝽,直立型叶片茸毛感筛豆龟蝽；紧贴型叶柄茸毛抗筛豆龟蝽,直立型叶柄茸毛感筛豆龟蝽,且叶片与叶柄相比,叶片的茸毛着生状态与筛豆龟蝽抗性的关联更为显著。
(3)重组自交系群体(NJRISX)中直立型叶片茸毛家系有85个,匍匐型叶片茸毛家系有91个,符合1：1的一对基因的基因型分离比,利用221个分子标记(218个SSR标记,3个形态标记)对叶片茸毛着生状态做连锁分析,将控制叶片茸毛着生状态的基因定位在H连锁群末端,与Satt317相距36.6 cM。
(4)以通山薄皮黄豆甲(T)×矮秆黄(A)组合,构建叶片茸毛着生状态的高代自交分离群体,选择到了目标区段杂合单株,通过田间表型鉴定和连续选择,得到7个单株衍生成的家系共353株,叶片茸毛匍匐为显性共252株,叶片茸毛直立为隐性共101株,卡平方测验符合3：1的一对基因的表型分离比,结合重组自交系群体(NJRISX)中1：1的基因型分离比,验证了叶片茸毛着生状态一对基因的遗传。利用H连锁群上9个SSR分子标记对构建的高代自交分离群体控制叶片茸毛着生状态的基因进行了定位,发现叶片茸毛着生状态基因与分子标记Satt434紧密连锁,将其定位在H连锁群上距Satt434约4.6 cM处。
Insect resistance of crop is often related to crop morphology, and the looking for suitable living environment and food of insects is often based on the feeling toward plant. Soybean [Glycine max (L.) Merr.] accessions with prostrate leaf blade pubescence show higher resistance than those with erect leaf blade pubescence. Classic genetics indicates that the trait of leaf blade pubescence status is controlled by several genes, however its phenotype also shows quantitative trait character which is easily affected by genotype, growing stage and environment conditions.
Globular stink bug (GSB) [Megacota cribraria (Fabricius)] can cause great damage to yield and quality of soybean. And it is the main pest in eastern coast of China in recent 20 years, showing increasing damage tendency. Breeding resistant varieties is an important measure to reduce the soybean loss caused by M. cribraria. Revealing genetic variation and exploring resistant genes in soybean varieties are priorities in the work of soybean insect-resistant breeding.
In this study, the researchers investigated the association between soybean leaf pubescence status and M. cribraria resistance of varieties, and compared leaf blade pubescence status with leaf petiole pubescence status to find out which association is more closely with M. cribraria resistance by using 400 varieties from 24 provinces of China. Furthermore, a gene controlling leaf blade pubescence status was verified and mapped by using recombinant inbred lines and high generation inbred separation lines.
The main results of this experiment are listed as follows:
(1) The coefficients of variation of leaf pubescence status and the resistance to M. cribraria were large in the selected soybean germplasm, and the relationship between varieties for GSB resistance and leaf pubescence status were detected to be significant correlation. There were more high resistant accessions and less high susceptible accessions in Southern China eco-region (including Middle and lower changjiang valleys, Central-south, Southwest plateau and Southwest trophic eco-regions) than that in Northeast China and HuangHuaiHai valleys in China, indicating resistance to M. cribraria in soybean being related with the geographic sources.
(2) Significant correlations were detected between leaf pubescence status and geographic sources damage rate. The correlation between resistance levels of varieties to M. cribraria and leaf pubescence status had also reached a significant level. Prostrate leaf pubescence was resistant, erect leaf pubescence was susceptible to GBS in field. Compare leaf blade pubescence status with leaf petiole pubescence status, leaf blade pubescence status showed a more significant correlation with M. cribraria resistant of soybean.
(3) There were 85 lines with erect leaf blade pubescence and 91 lines with prostrate leaf blade pubescence in recombined inbred lines (NJRISX). It fitted the expected genotype segregation ratio of 1:1. According to the linkage analysis between the genotype (including 218 SSR markers and 3 morphological markers) and pheonotype informations, the gene cotrolling leaf blade pubescence was located at the end part of H linkage group,36.6 cM away from SSR marker Satt317.
(4) Some high generation inbred seperation lines were constructed. The population is selected from the lines derived from the cross of two soybean varieties T and A. Firstly, four single plants with heterozygous marker genotypes at the targeted genome regions were selected, and other genome regions of these plants remained homozygous. By subsequently genotypic and field phenotypic investigation, seven different heterozygous lines of the leaf blade pubescence status were developed. The pooled lines contained 353 plants, in which 252 plants showed dominant phenotype (prostrate leaf blade pubescence) and 101 plants showed recessive phenotype (erect leaf blade pubescence). The phenotype segregation pattern fitted 3:1 expected ratio. Based on the 1:1 genotype segregation ratio of recombined inbred lines (NJRISX), the conclusion that this character was controlled by a single major gene was verified. Using the high generation inbred seperation lines and 9 SSR markers to fine map the targeted gene, the result showed the targeted gene was tightly linked with two SSR markers Satt434 and 12-1364a, and located the gene around the marker Satt434 about 4.6 cM.
本研究利用来源于全国24个省份的400份大豆地方品种研究大豆叶茸毛着生状态与筛豆龟蝽抗性的关系,比较叶片和叶柄茸毛着生状态与筛豆龟蝽抗性的关联；进一步通过重组自交系群体和构建的高代自交分离群体,对控制叶片茸毛着生状态的基因进行定位和验证,为叶片茸毛着生状态基因的育种利用及其与抗虫性的相互作用等研究提供指导。本研究的主要结果如下：
(1)大豆种质资源品种间对筛豆龟蝽抗性存在较大的变异,筛豆龟蝽的抗性等级与生态区间也存在显著的相关,东北、黄淮海生态区的高感品种的频率高于长江中下游,中南,西南高原和华南热带等南方生态区,而长江中下游,中南,西南高原和华南热带等南方生态区高抗品种的频率高于东北、黄淮海生态区。
(2)叶茸毛着生状态与筛豆龟蝽的危害率两者间存在极显著关联。匍匐型叶片茸毛抗筛豆龟蝽,直立型叶片茸毛感筛豆龟蝽；紧贴型叶柄茸毛抗筛豆龟蝽,直立型叶柄茸毛感筛豆龟蝽,且叶片与叶柄相比,叶片的茸毛着生状态与筛豆龟蝽抗性的关联更为显著。
(3)重组自交系群体(NJRISX)中直立型叶片茸毛家系有85个,匍匐型叶片茸毛家系有91个,符合1：1的一对基因的基因型分离比,利用221个分子标记(218个SSR标记,3个形态标记)对叶片茸毛着生状态做连锁分析,将控制叶片茸毛着生状态的基因定位在H连锁群末端,与Satt317相距36.6 cM。
(4)以通山薄皮黄豆甲(T)×矮秆黄(A)组合,构建叶片茸毛着生状态的高代自交分离群体,选择到了目标区段杂合单株,通过田间表型鉴定和连续选择,得到7个单株衍生成的家系共353株,叶片茸毛匍匐为显性共252株,叶片茸毛直立为隐性共101株,卡平方测验符合3：1的一对基因的表型分离比,结合重组自交系群体(NJRISX)中1：1的基因型分离比,验证了叶片茸毛着生状态一对基因的遗传。利用H连锁群上9个SSR分子标记对构建的高代自交分离群体控制叶片茸毛着生状态的基因进行了定位,发现叶片茸毛着生状态基因与分子标记Satt434紧密连锁,将其定位在H连锁群上距Satt434约4.6 cM处。
Insect resistance of crop is often related to crop morphology, and the looking for suitable living environment and food of insects is often based on the feeling toward plant. Soybean [Glycine max (L.) Merr.] accessions with prostrate leaf blade pubescence show higher resistance than those with erect leaf blade pubescence. Classic genetics indicates that the trait of leaf blade pubescence status is controlled by several genes, however its phenotype also shows quantitative trait character which is easily affected by genotype, growing stage and environment conditions.
Globular stink bug (GSB) [Megacota cribraria (Fabricius)] can cause great damage to yield and quality of soybean. And it is the main pest in eastern coast of China in recent 20 years, showing increasing damage tendency. Breeding resistant varieties is an important measure to reduce the soybean loss caused by M. cribraria. Revealing genetic variation and exploring resistant genes in soybean varieties are priorities in the work of soybean insect-resistant breeding.
In this study, the researchers investigated the association between soybean leaf pubescence status and M. cribraria resistance of varieties, and compared leaf blade pubescence status with leaf petiole pubescence status to find out which association is more closely with M. cribraria resistance by using 400 varieties from 24 provinces of China. Furthermore, a gene controlling leaf blade pubescence status was verified and mapped by using recombinant inbred lines and high generation inbred separation lines.
The main results of this experiment are listed as follows:
(1) The coefficients of variation of leaf pubescence status and the resistance to M. cribraria were large in the selected soybean germplasm, and the relationship between varieties for GSB resistance and leaf pubescence status were detected to be significant correlation. There were more high resistant accessions and less high susceptible accessions in Southern China eco-region (including Middle and lower changjiang valleys, Central-south, Southwest plateau and Southwest trophic eco-regions) than that in Northeast China and HuangHuaiHai valleys in China, indicating resistance to M. cribraria in soybean being related with the geographic sources.
(2) Significant correlations were detected between leaf pubescence status and geographic sources damage rate. The correlation between resistance levels of varieties to M. cribraria and leaf pubescence status had also reached a significant level. Prostrate leaf pubescence was resistant, erect leaf pubescence was susceptible to GBS in field. Compare leaf blade pubescence status with leaf petiole pubescence status, leaf blade pubescence status showed a more significant correlation with M. cribraria resistant of soybean.
(3) There were 85 lines with erect leaf blade pubescence and 91 lines with prostrate leaf blade pubescence in recombined inbred lines (NJRISX). It fitted the expected genotype segregation ratio of 1:1. According to the linkage analysis between the genotype (including 218 SSR markers and 3 morphological markers) and pheonotype informations, the gene cotrolling leaf blade pubescence was located at the end part of H linkage group,36.6 cM away from SSR marker Satt317.
(4) Some high generation inbred seperation lines were constructed. The population is selected from the lines derived from the cross of two soybean varieties T and A. Firstly, four single plants with heterozygous marker genotypes at the targeted genome regions were selected, and other genome regions of these plants remained homozygous. By subsequently genotypic and field phenotypic investigation, seven different heterozygous lines of the leaf blade pubescence status were developed. The pooled lines contained 353 plants, in which 252 plants showed dominant phenotype (prostrate leaf blade pubescence) and 101 plants showed recessive phenotype (erect leaf blade pubescence). The phenotype segregation pattern fitted 3:1 expected ratio. Based on the 1:1 genotype segregation ratio of recombined inbred lines (NJRISX), the conclusion that this character was controlled by a single major gene was verified. Using the high generation inbred seperation lines and 9 SSR markers to fine map the targeted gene, the result showed the targeted gene was tightly linked with two SSR markers Satt434 and 12-1364a, and located the gene around the marker Satt434 about 4.6 cM.
引文
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