萝卜肉质根根重性状遗传标记分析与膨胀素基因家族的克隆
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摘要
萝卜(Raphanus sativus L.)是起源于我国的一种重要世界性蔬菜,栽培面积较大,以膨大的肉质根为主要食用部位。本文利用植物数量性状主基因+多基因混合遗传模型多世代联合分析法,探讨了萝卜肉质根根重的遗传规律；采用基因型代表群分析法(GRA)对肉质根根重性状进行RAPD标记分析；同时克隆了膨胀素基因家族,并初步研究其在萝卜不同发育时期中的表达特征。
以肉质根大小差异显著的萝卜高代自交系Nau-WH05、Nau-XWFDH05、Nau-LH05和Nau-LH06配制杂交组合,利用植物数量性状主基因+多基因混合遗传模型多世代联合分析方法,探讨萝卜肉质根根重的遗传规律。结果表明：组合Nau-WH05×Nau-LH05根重遗传符合两对加性-显性-上位性主基因+加性-显性-上位性多基因混合遗传模型；组合Nau-XWFDH05×Nau-LH06符合一对加性-显性主基因+加性-显性上位性多基因遗传模型。表明萝卜肉质根根重是由主基因和多基因控制的数量性状。
以高代自交系NC、CDG、BYC、CB和WH、LHZ及其F1、F2群体为材料,采用基因型代表群分析法(GRA)对萝卜肉质根根重性状进行RAPD标记分析,在近缘野生种LHZ中扩增出一条特异条带NAURP7941200。WH×LHZ的F2群体分析表明,该标记可能与肉质根根重性状相关。
利用简并引物扩增和电子克隆的方法,从萝卜高根冠比自交系NAU-LVYH06中获得了2个膨胀素基因片段和13个膨胀素基因。其中仅2个属于β-亚族,其余属于α-亚族。在所获得的膨胀素基因DNA序列中,均存在内含子,但内含子的数目和长度存在差异；内含子插入模式分析结果表明：7个α-亚族的膨胀素均含两个内含子,插入位置相同,为C和E插入位点；2个β-亚族均含有三个内含子,插入位置相同,为C、E和F位点。共检测到三个内含子插入位点(C、E和F),未检测到插入位点A、B和D。内含子C和E为所有基因共有,长度差异较大(79bp-604bp)。内含子F为两个p-亚族所具有,长度变化较小(243bp-316bp)。推导氨基酸序列均存在信号肽,等电点和分子量差别较小。
Radish(Raphanus sativus L.) is a world vegetable, originated from China. It has been widely cultivated and plays a very important role in vegetable production in our country. The method of major gene plus poly genes mixed inheritance model was used to analyze the genetics of tap root weight. RAPD markers were scanned based on the strategy of Genoty-pic Representation Analysis (GRA). We also cloned the expansin gene family and analyzed their expression profiles and putative functions in different development stages.
Genetic analysis was conducted on tap root weight with multiple generations. P1, P2, F1 and F2 were derived from across between big tap root type Nau-WH05, Nau-XWFDH05 and small tap root type Nau-LH05, Nau-LH06, using the method of major gene plus polygenes mixed inheritance model inquantitative traits of plants. It could be concluded that the optimum model of tap root weight were two additive-dominance-epistatic major gene plus adding-dominance-epistatic polygenes in cross Nau-WH05×Nau-LH05 and one adding-dominance major gene plus adding-dominance-epistatic polygenes in cross Nau-XWFDH05×Nau-LHZ06. The tap root weight was quantitative trait and controlled by major gene(s) plus polygenes in radish.
With NC、CDG、BYC、CB and WH、LHZ、F1、F2 population as materials, The trait of tap root weight was researched using GRA strategy and RAPD technology. A RAPD marker NAURP7941200 was screened. Based on ananysis of the F2 population derived from WH×LHZ, the maker may linked with tap root weight.
Two expansin gene fragments and thirteen expansin genes were obtained by degenerate primers PCR and insilico clonnig. two wereβ-expansin and the others wereα-expansin. The intorns were existed in nine expansin gene genome sequences, but the numbers and length were different. In this study, three intorns, C, E and F, could be observed. It was shown that position C and E (79bp-604bp) are present in both a andβ-expansin genes. Positions F (243bp-316bp) were only present inβ-expansin, suggesting that they might occurringβ-expansin shortly afterα/βsubgroup split. All the deduced amino acids contained a single peptide, PI and MW were approximately.
以肉质根大小差异显著的萝卜高代自交系Nau-WH05、Nau-XWFDH05、Nau-LH05和Nau-LH06配制杂交组合,利用植物数量性状主基因+多基因混合遗传模型多世代联合分析方法,探讨萝卜肉质根根重的遗传规律。结果表明：组合Nau-WH05×Nau-LH05根重遗传符合两对加性-显性-上位性主基因+加性-显性-上位性多基因混合遗传模型；组合Nau-XWFDH05×Nau-LH06符合一对加性-显性主基因+加性-显性上位性多基因遗传模型。表明萝卜肉质根根重是由主基因和多基因控制的数量性状。
以高代自交系NC、CDG、BYC、CB和WH、LHZ及其F1、F2群体为材料,采用基因型代表群分析法(GRA)对萝卜肉质根根重性状进行RAPD标记分析,在近缘野生种LHZ中扩增出一条特异条带NAURP7941200。WH×LHZ的F2群体分析表明,该标记可能与肉质根根重性状相关。
利用简并引物扩增和电子克隆的方法,从萝卜高根冠比自交系NAU-LVYH06中获得了2个膨胀素基因片段和13个膨胀素基因。其中仅2个属于β-亚族,其余属于α-亚族。在所获得的膨胀素基因DNA序列中,均存在内含子,但内含子的数目和长度存在差异；内含子插入模式分析结果表明：7个α-亚族的膨胀素均含两个内含子,插入位置相同,为C和E插入位点；2个β-亚族均含有三个内含子,插入位置相同,为C、E和F位点。共检测到三个内含子插入位点(C、E和F),未检测到插入位点A、B和D。内含子C和E为所有基因共有,长度差异较大(79bp-604bp)。内含子F为两个p-亚族所具有,长度变化较小(243bp-316bp)。推导氨基酸序列均存在信号肽,等电点和分子量差别较小。
Radish(Raphanus sativus L.) is a world vegetable, originated from China. It has been widely cultivated and plays a very important role in vegetable production in our country. The method of major gene plus poly genes mixed inheritance model was used to analyze the genetics of tap root weight. RAPD markers were scanned based on the strategy of Genoty-pic Representation Analysis (GRA). We also cloned the expansin gene family and analyzed their expression profiles and putative functions in different development stages.
Genetic analysis was conducted on tap root weight with multiple generations. P1, P2, F1 and F2 were derived from across between big tap root type Nau-WH05, Nau-XWFDH05 and small tap root type Nau-LH05, Nau-LH06, using the method of major gene plus polygenes mixed inheritance model inquantitative traits of plants. It could be concluded that the optimum model of tap root weight were two additive-dominance-epistatic major gene plus adding-dominance-epistatic polygenes in cross Nau-WH05×Nau-LH05 and one adding-dominance major gene plus adding-dominance-epistatic polygenes in cross Nau-XWFDH05×Nau-LHZ06. The tap root weight was quantitative trait and controlled by major gene(s) plus polygenes in radish.
With NC、CDG、BYC、CB and WH、LHZ、F1、F2 population as materials, The trait of tap root weight was researched using GRA strategy and RAPD technology. A RAPD marker NAURP7941200 was screened. Based on ananysis of the F2 population derived from WH×LHZ, the maker may linked with tap root weight.
Two expansin gene fragments and thirteen expansin genes were obtained by degenerate primers PCR and insilico clonnig. two wereβ-expansin and the others wereα-expansin. The intorns were existed in nine expansin gene genome sequences, but the numbers and length were different. In this study, three intorns, C, E and F, could be observed. It was shown that position C and E (79bp-604bp) are present in both a andβ-expansin genes. Positions F (243bp-316bp) were only present inβ-expansin, suggesting that they might occurringβ-expansin shortly afterα/βsubgroup split. All the deduced amino acids contained a single peptide, PI and MW were approximately.
引文
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