彩色马铃薯新品系细胞遗传学观测及SSR分析

doi:10.7668/hbnxb.20193007

摘要/Abstract

摘要：

为了明确6个彩色马铃薯新品系NNCS-1、NNCS-5、NNCS-6、NNCS-7、NNCS-33和NNCS-37在细胞遗传学和DNA分子水平上的遗传差异,以马铃薯材料MIN-021为对照,采用染色体制片法和SSR标记技术,对其花粉育性、花粉母细胞减数分裂中期 Ⅰ 染色体构型和SSR进行了分析。结果表明,6个彩色马铃薯新品系的花粉育性依次分别为44.00%,66.78%,74.44%,60.70%,78.10%,80.22%,其染色体配对构型依次为8.45Ⅰ+6.09Ⅱ+7.35Ⅲ+1.33Ⅳ、6.91Ⅰ+7.13Ⅱ+5.73Ⅲ+2.41Ⅳ、5.19Ⅰ+9.16Ⅱ+4.51Ⅲ+2.74Ⅳ、6.55Ⅰ+8.35Ⅱ+4.97Ⅲ+2.46Ⅳ、4.02Ⅰ+10.15Ⅱ+3.40Ⅲ+3.37Ⅳ和3.31Ⅰ+10.42Ⅱ+2.99Ⅲ+3.72Ⅳ,表明在细胞学水平上各品系间有一定的差异。利用筛选出的10对SSR适宜引物,对各新品系及对照材料的基因组DNA进行扩增获得SSR多态性位点151个,多态性位点比率占86.78%,各材料间遗传差异较大。用筛选出的特异性引物C42建立了1张能识别出6个彩色马铃薯新品系及对照的SSR指纹图。以平均遗传距离(GD值)0.62为基准,将7个材料分为3类:新品系NNCS-37、NNCS-6、NNCS-7、NNCS-5和对照(MIN-021)为一类;新品系NNCS-33和NNCS-1各单独为一类。

关键词: 彩色马铃薯新品系, 细胞遗传学, SSR分析

Abstract:

In order to clarify the genetic differences of the six new-colored potato strains NNCS-1,NNCS-5,NNCS-6,NNCS-7,NNCS-33 and NNCS-37 at the cytogenetic and DNA molecular level,this experiment used material MIN-021 as control,and the regular staining and slides preparing and SSR marker technology were used to analyze the pollen fertility,chromosome configuration at meiosis metaphase Ⅰ of pollen mother cells and SSR polymorphism of the above six varieties.The results shaved that the pollen fertility of the six new-colored potato strains were 44.00%,66.78%,74.44%,60.70%,78.10% and 80.22%,respectively.And the chromosome pairing configurations of these test materials were 8.45Ⅰ+6.09Ⅱ+7.35Ⅲ+1.33Ⅳ,6.91Ⅰ+7.13Ⅱ+5.73Ⅲ+2.41Ⅳ,5.19Ⅰ+9.16Ⅱ+4.51Ⅲ+2.74Ⅳ,6.55Ⅰ+8.35Ⅱ+4.97Ⅲ+2.46Ⅳ,4.02Ⅰ+10.15Ⅱ+3.40Ⅲ+3.37Ⅳ and 3.31Ⅰ+10.42Ⅱ+2.99Ⅲ+3.72Ⅳ,respectively,indicating that there were certain differences among the strains at the cytological level.Ten suitable SSR primers were selected to amplify the genomic DNA of each new strain and control material,and 151 SSR polymorphic sites were obtained,accounting for 86.78% of the total number of polymorphic loci.The genetic differences among the materials were revealed.In addition,a SSR fingerprint was amplified with the selected specific primers C42,which could identify six new-colored potato strains and control.Based on the average genetic distance(GD=0.62),the seven materials were divided into three categories: the new strain NNCS-37,NNCS-6,NNCS-7,NNCS-5 and the control(MIN-021)belong to one categorie.The new strains NNCS-33 and NNCS-1 were separately classified as a categorie.

Key words: New colored potato strain, Cytogenetics, SSR analysis

岳东, 于卓, 于肖夏, 李佳奇, 李景伟, 杨东升, 张海龙, 李志聪. 彩色马铃薯新品系细胞遗传学观测及SSR分析[J]. 华北农学报, 2022, 37(6): 108-114. doi: 10.7668/hbnxb.20193007.

YUE Dong, YU Zhuo, YU Xiaoxia, LI Jiaqi, LI Jingwei, YANG Dongsheng, ZHANG Hailong, LI Zhicong. Cytological Observation and SSR Analysis of New Colored Potato Strains[J]. Acta Agriculturae Boreali-Sinica, 2022, 37(6): 108-114. doi: 10.7668/hbnxb.20193007.

http://www.hbnxb.net/CN/Y2022/V37/I6/108

图/表 10

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引物名称 Primer name	正向引物(5'—3') Forward primer(5'—3')	反向引物(5'—3') Reversed primer(5'—3')
S7	GACTGGCTGACCCTGAACTC	GACAAAATTACAGGAACTGCAAA
S25	GCGAATGACAGGACAAGAGG	TGCCACTGCTACCATAACCA
S153	TATGTTCCACGCCATTTCAG	ACGGAAACTCATCGTGCATT
S174	TGAGGGTTTTCAGAAAGGGA	CATCCTTGCAACAACCTCCT
S189	CCTTGTAGTACAGCAGTGGTC	TCCGCCAAGACTGATGCA
C42	AGGCAAACCCCGTAAATCTT	TCTTCCTCACTCGTCCCACT
C60	ACTTCTGCATCTGGTGAAGC	GGTCGGATTCCCAGGTTG
STI014	AGAAACTGAGTTGTGTTGGGA	TCAACAGTCTCAGAAAACCCTCT
STM1016	TTCTGATTTCATGCATGTTTCC	ATGCTTGCCATGTGATGTGT
STM2022	GCGTCAGCGATTTCAGTACTA	TTCAGTCAACTCCTGTTGCG

引物名称 Primer name	正向引物(5'—3') Forward primer(5'—3')	反向引物(5'—3') Reversed primer(5'—3')
S7	GACTGGCTGACCCTGAACTC	GACAAAATTACAGGAACTGCAAA
S25	GCGAATGACAGGACAAGAGG	TGCCACTGCTACCATAACCA
S153	TATGTTCCACGCCATTTCAG	ACGGAAACTCATCGTGCATT
S174	TGAGGGTTTTCAGAAAGGGA	CATCCTTGCAACAACCTCCT
S189	CCTTGTAGTACAGCAGTGGTC	TCCGCCAAGACTGATGCA
C42	AGGCAAACCCCGTAAATCTT	TCTTCCTCACTCGTCCCACT
C60	ACTTCTGCATCTGGTGAAGC	GGTCGGATTCCCAGGTTG
STI014	AGAAACTGAGTTGTGTTGGGA	TCAACAGTCTCAGAAAACCCTCT
STM1016	TTCTGATTTCATGCATGTTTCC	ATGCTTGCCATGTGATGTGT
STM2022	GCGTCAGCGATTTCAGTACTA	TTCAGTCAACTCCTGTTGCG

材料 Materials	可育花粉粒数目 Pollen grain fertility number	总的花粉粒数目 Total pollen grain number	花粉可育率 /% Pollen grain fertility rate
NNCS-33	3 519	4 506	78.10
NNCS-37	4 103	5 115	80.22
NNCS-6	3 381	4 542	74.44
NNCS-7	3 651	6 015	60.70
NNCS-5	4 716	7 062	66.78
NNCS-1	2 506	5 696	44.00
MIN-021(CK)	3 869	5 144	75.21

材料 Materials	可育花粉粒数目 Pollen grain fertility number	总的花粉粒数目 Total pollen grain number	花粉可育率 /% Pollen grain fertility rate
NNCS-33	3 519	4 506	78.10
NNCS-37	4 103	5 115	80.22
NNCS-6	3 381	4 542	74.44
NNCS-7	3 651	6 015	60.70
NNCS-5	4 716	7 062	66.78
NNCS-1	2 506	5 696	44.00
MIN-021(CK)	3 869	5 144	75.21

引物名称 Primer name	多态性条带 Number of polymorphic loci	扩增条带总数 Number of amplified loci	多态性比率 /% Percentage of polymorphic loci
S7	12	14	85.71
S25	13	16	81.25
S153	16	18	88.89
S174	13	15	86.67
S189	16	17	94.12
C42	23	24	95.83
C60	13	17	76.47
STI014	13	16	81.25
STM2022	15	18	83.33
STM1016	17	19	89.47
总计Total	151	174	86.78

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