3个不同类型尖叶胡枝子遗传多样性研究
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摘要
以尖叶胡枝子(Lespedeza hedysaroides (Pall.) Kitag.)的3个不同类型为材料，从形态、生化、细胞和分子4个标记水平研究其遗传多样性，主要结果如下：
1.3个不同类型尖叶胡枝子具有丰富的表型多样性，表现出显著性差异水平的指标包括株高、基部茎粗、叶片、种子、粗枝重和中枝重等19个性状；相关分析中呈正相关关系的有14对性状，呈负相关关系的有16对性状；第一主成分主要指标按作用大小排列为绝对株高、自然株高、二级分枝、基部茎粗、花梗长和一级分枝等20个性状，代表了70%的全部信息。
2.利用SDS-PAGE获得的盐溶蛋白图谱和贮藏蛋白图谱能很好的区分3个不同类型的尖叶胡枝子，蛋白分子量介于12.43～154.53kDa之间。盐溶蛋白总谱带数为19条，多态率为68.42%；贮藏蛋白总谱带数为28条，多态率为32.14%。
3.3个不同类型尖叶胡枝子的染色体数目相同（2n=2x=20），为二倍体。普通型尖叶胡枝子的核型公式为2n=2x=14m+4sm+2T(SAT)，核型为2B型，核型不对称系数为60.63%；浓绿型尖叶胡枝子的核型公式与普通型相同，核型为2A型，核型不对称系数为62.01%；高杆型尖叶胡枝子的核型公式为2n=2x=12m+4sm+4T(SAT)，核型为2B型，核型不对称系数为64.16%。
4.9个ISSR引物共扩增出109条清晰稳定的条带，每个引物平均多态性为88.91%。3个不同类型尖叶胡枝子的有效等位基因数为1.6996，基因多样性指数为0.3908，Shannon指数为0.5601；聚类结果为浓绿型和高杆型尖叶胡枝子聚为一类，普通型尖叶胡枝子单独聚为一类。表型性状和蛋白多样性聚类分析表明，普通型和浓绿型尖叶胡枝子聚为一类，高杆型尖叶胡枝子单独聚为一类。3个不同类型尖叶胡枝子之间存在明显差异，其中高杆型和普通型尖叶胡枝子之间的差异较大。
The research studied the genetic diversity from morphology, protein, chromosome and moleculeslevels in three types of Lespedeza hedysaroides (Pall.) Kitag.. Main results were as followed:
1. Three types of Lespedeza hedysaroides were rich of morphological diversity. The ANOVAanalysis results showed that, plant height, basal stem diameter, leaf, seed, thick stem weight and middlestem weight et al. total19traits showed a significant difference in three types of Lespedezahedysaroides. Correlation analysis showed14pairs of traits were significantly positive correlation and16pairs of traits were significant negative correlation. The main indicators of the first principalcomponent, which were arrayed as effect degree including the plant height, sublevel branches, basestem diameter, footstalk length, primary branches, et al. total20indexs delegating70%of the totalinformation.
2. Salt soluble proteins and seed storage proteins in the common-type and deep green-type ofLespedeza hedysaroides were different from the tall-type. The protein molecular weights ranged from12.43to154.53kDa.6bands were shared among the three types and13bands were specific andpolymorphism accounted for68.42%of total19salt soluble protein bands. In the total seed storageproteins were19bands shared and9bands were specific and polymorphism accounted here for32.14%of the total28bands.
3. The number of chromosomes were the same among3types of Lespedeza hedysaroides, whichwas2n=2x=20, belonged to diploid. The karyotype formula was2n=2x=14m+4sm+2T (SAT) of thecommon-type Lespedeza hedysaroides, which was2B type; the karyotype asymmetric coefficient was60.63%. The deep green-type’s karyotype formula was the same to the common-type, but was2A type,and the karyotype asymmetric coefficient was62.01%; the tall-type Lespedeza hedysaroides karyotypeformula was2n=2x=12m+4sm+4T (SAT), belonged to2B type, the karyotype asymmetric coefficientwas64.16%.
4. Nine primers of ISSR were utilized and total109bands were obtained by amplication reaction,each primer showed average88.91%of polymorphism. The effective number of alleles was1.6996, theindex of Nei’s genetic diversity was0.3908and the Shannon’s information index was0.5601of threetypes of Lespedeza hedysaroides; ISSR clustering showed that the deep green-type and tall-type wereclassified as one group; the common-type was another. The dendrogram of phenotype and proteinsdiversity suggested that the common-type and deep green-type were clustered as one group, the tall-typewere classified as a different group. There were obvious differences among three types of Lespedezahedysaroides and the difference between the tall-type and the common-type Lespedeza hedysaroideswas significant.
1.3个不同类型尖叶胡枝子具有丰富的表型多样性，表现出显著性差异水平的指标包括株高、基部茎粗、叶片、种子、粗枝重和中枝重等19个性状；相关分析中呈正相关关系的有14对性状，呈负相关关系的有16对性状；第一主成分主要指标按作用大小排列为绝对株高、自然株高、二级分枝、基部茎粗、花梗长和一级分枝等20个性状，代表了70%的全部信息。
2.利用SDS-PAGE获得的盐溶蛋白图谱和贮藏蛋白图谱能很好的区分3个不同类型的尖叶胡枝子，蛋白分子量介于12.43～154.53kDa之间。盐溶蛋白总谱带数为19条，多态率为68.42%；贮藏蛋白总谱带数为28条，多态率为32.14%。
3.3个不同类型尖叶胡枝子的染色体数目相同（2n=2x=20），为二倍体。普通型尖叶胡枝子的核型公式为2n=2x=14m+4sm+2T(SAT)，核型为2B型，核型不对称系数为60.63%；浓绿型尖叶胡枝子的核型公式与普通型相同，核型为2A型，核型不对称系数为62.01%；高杆型尖叶胡枝子的核型公式为2n=2x=12m+4sm+4T(SAT)，核型为2B型，核型不对称系数为64.16%。
4.9个ISSR引物共扩增出109条清晰稳定的条带，每个引物平均多态性为88.91%。3个不同类型尖叶胡枝子的有效等位基因数为1.6996，基因多样性指数为0.3908，Shannon指数为0.5601；聚类结果为浓绿型和高杆型尖叶胡枝子聚为一类，普通型尖叶胡枝子单独聚为一类。表型性状和蛋白多样性聚类分析表明，普通型和浓绿型尖叶胡枝子聚为一类，高杆型尖叶胡枝子单独聚为一类。3个不同类型尖叶胡枝子之间存在明显差异，其中高杆型和普通型尖叶胡枝子之间的差异较大。
The research studied the genetic diversity from morphology, protein, chromosome and moleculeslevels in three types of Lespedeza hedysaroides (Pall.) Kitag.. Main results were as followed:
1. Three types of Lespedeza hedysaroides were rich of morphological diversity. The ANOVAanalysis results showed that, plant height, basal stem diameter, leaf, seed, thick stem weight and middlestem weight et al. total19traits showed a significant difference in three types of Lespedezahedysaroides. Correlation analysis showed14pairs of traits were significantly positive correlation and16pairs of traits were significant negative correlation. The main indicators of the first principalcomponent, which were arrayed as effect degree including the plant height, sublevel branches, basestem diameter, footstalk length, primary branches, et al. total20indexs delegating70%of the totalinformation.
2. Salt soluble proteins and seed storage proteins in the common-type and deep green-type ofLespedeza hedysaroides were different from the tall-type. The protein molecular weights ranged from12.43to154.53kDa.6bands were shared among the three types and13bands were specific andpolymorphism accounted for68.42%of total19salt soluble protein bands. In the total seed storageproteins were19bands shared and9bands were specific and polymorphism accounted here for32.14%of the total28bands.
3. The number of chromosomes were the same among3types of Lespedeza hedysaroides, whichwas2n=2x=20, belonged to diploid. The karyotype formula was2n=2x=14m+4sm+2T (SAT) of thecommon-type Lespedeza hedysaroides, which was2B type; the karyotype asymmetric coefficient was60.63%. The deep green-type’s karyotype formula was the same to the common-type, but was2A type,and the karyotype asymmetric coefficient was62.01%; the tall-type Lespedeza hedysaroides karyotypeformula was2n=2x=12m+4sm+4T (SAT), belonged to2B type, the karyotype asymmetric coefficientwas64.16%.
4. Nine primers of ISSR were utilized and total109bands were obtained by amplication reaction,each primer showed average88.91%of polymorphism. The effective number of alleles was1.6996, theindex of Nei’s genetic diversity was0.3908and the Shannon’s information index was0.5601of threetypes of Lespedeza hedysaroides; ISSR clustering showed that the deep green-type and tall-type wereclassified as one group; the common-type was another. The dendrogram of phenotype and proteinsdiversity suggested that the common-type and deep green-type were clustered as one group, the tall-typewere classified as a different group. There were obvious differences among three types of Lespedezahedysaroides and the difference between the tall-type and the common-type Lespedeza hedysaroideswas significant.
引文
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