不同小麦品种穗发芽特性的鉴定及TaRHA2b基因序列差异分析
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摘要
为了开发新的抗穗发芽分子标记,以50个发芽指数具有显著差异的小麦品种为材料,进行穗发芽特性的鉴定,同时对这50个小麦品种的RING finger E3连接酶基因TaRHA2b的核苷酸序列进行测定,然后进行比对分析。结果表明,50个小麦品种的抗穗发芽能力有明显差异;不同小麦品种间TaRHA2b基因序列的长度变化不大,但是存在6个单核苷酸多态性位点,其中第47位碱基的差异造成EcoICRⅠ/Ecl136Ⅱ和SacⅠ/SstⅠ这2个酶切位点的差异。
In order to develop a new pre-harvest sprouting resistant marker,fifty wheat varieties with significantly different germination index were selected as experimental materials,and their germination characteristics were studied,the sequences of RING finger E3 ligase gene TaRHA2b of these fifty wheat varieties were determined and compared. The results showed that the resistance to pre-harvest sprouting was obviously different among the fifty wheat varieties; the length of the TaRHA2b gene sequences varied little among different wheat varieties,but there were six single nucleotide polymorphisms,the difference of 47 th base resulted in differences in the two cleavage sites of EcoICRⅠ/Ecl136Ⅱ and SacⅠ/SstⅠ.
In order to develop a new pre-harvest sprouting resistant marker,fifty wheat varieties with significantly different germination index were selected as experimental materials,and their germination characteristics were studied,the sequences of RING finger E3 ligase gene TaRHA2b of these fifty wheat varieties were determined and compared. The results showed that the resistance to pre-harvest sprouting was obviously different among the fifty wheat varieties; the length of the TaRHA2b gene sequences varied little among different wheat varieties,but there were six single nucleotide polymorphisms,the difference of 47 th base resulted in differences in the two cleavage sites of EcoICRⅠ/Ecl136Ⅱ and SacⅠ/SstⅠ.
引文
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[6]吕军朋. RING finger类基因功能研究及抗穗发芽转基因小麦的创制[D].郑州:河南农业大学,2011.
[7] STONE S L,HAUKSDTTIR H,TROY A,et al. Functional analysis of the RING-type ubiquitin ligase family of Arabidopsis[J]. Plant Physiology,2005,137(1):13-30.
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[9] CHEN D H,RONALD P C. A rapid DNA minipreparation method suitable for AFLP and other PCR applications[J]. Plant Molecular Biology Reporter,1999,17(1):53-57.
[10] GUBLER F,MILLAR A A,JACOBSEN J V. Dormancy release,ABA and pre-harvest sprouting[J]. Curr Opin Plant Biol,2005,8(2):183-187.
[11] MCKIBBIN R S,WILKINSON M D,BAILEY P C,et al.Transcripts of Vp-1 homeologues are misspliced in modern wheat and ancestral species[J]. Proc Natl Acad Sci USA,2002,99(15):10203-10208.
[12]高永峰,刘继恺,范晶,等.水稻穗发芽调控基因OsVP1的RNA干涉载体构建及遗传转化研究[J].中国农业科学,2010,43(7):1321-1327.
[13] YANG Y,ZHAO X L,XIA L Q,et al. Development and validation of a Viviparous-1 STS marker for pre-harvest sprouting tolerance in Chinese wheats[J]. Theoretical&Applied Genetics,2007,115(7):971-980.
[14] ZHANG Y,XIA X,HE Z. The seed dormancy allele Ta Sdr-A1a associated with pre-harvest sprouting tolerance is mainly present in Chinese wheat landraces[J].Theoretical&Applied Genetics,2017,130(1):81-89.
[2]任立世,刘进英,杨燕. 107份小麦历史品种抗穗发芽基因型的检测[J].麦类作物学报,2015,35(6):752-758.
[3] BU Q Y,LI H M,ZHAO Q Z,et al. The Arabidopsis RING finger E3 ligase RHA2a is a novel positive regulator of abscisic acid signaling during seed germination and early seedling development[J]. Plant Physiology,2009,150(1):463-481.
[4] BU Q,JIANG H,LI C B,et al. Role of the Arabidopsis thaliana NAC transcription factors ANAC019 and ANAC055 in regulating jasmonic acid-signaled defense responses[J]. Cell Res,2008,18(7):756-767.
[5] LI H,JIANG H,BU Q,et al. The Arabidopsis RING finger E3 ligase RHA2b acts additively with RHA2a in regulating abscisic acid signaling and drought response[J].Plant Physiol,2011,156(2):550-563.
[6]吕军朋. RING finger类基因功能研究及抗穗发芽转基因小麦的创制[D].郑州:河南农业大学,2011.
[7] STONE S L,HAUKSDTTIR H,TROY A,et al. Functional analysis of the RING-type ubiquitin ligase family of Arabidopsis[J]. Plant Physiology,2005,137(1):13-30.
[8] VIERSTRA R D. The ubiquitin-26S proteasome system at the nexus of plant biology[J]. Nature Reviews Molecular Cell Biology,2009,10(6):385-397.
[9] CHEN D H,RONALD P C. A rapid DNA minipreparation method suitable for AFLP and other PCR applications[J]. Plant Molecular Biology Reporter,1999,17(1):53-57.
[10] GUBLER F,MILLAR A A,JACOBSEN J V. Dormancy release,ABA and pre-harvest sprouting[J]. Curr Opin Plant Biol,2005,8(2):183-187.
[11] MCKIBBIN R S,WILKINSON M D,BAILEY P C,et al.Transcripts of Vp-1 homeologues are misspliced in modern wheat and ancestral species[J]. Proc Natl Acad Sci USA,2002,99(15):10203-10208.
[12]高永峰,刘继恺,范晶,等.水稻穗发芽调控基因OsVP1的RNA干涉载体构建及遗传转化研究[J].中国农业科学,2010,43(7):1321-1327.
[13] YANG Y,ZHAO X L,XIA L Q,et al. Development and validation of a Viviparous-1 STS marker for pre-harvest sprouting tolerance in Chinese wheats[J]. Theoretical&Applied Genetics,2007,115(7):971-980.
[14] ZHANG Y,XIA X,HE Z. The seed dormancy allele Ta Sdr-A1a associated with pre-harvest sprouting tolerance is mainly present in Chinese wheat landraces[J].Theoretical&Applied Genetics,2017,130(1):81-89.