华北农学报
華北農學報
화북농학보
ACTA AGRICULTURAE BOREALI-SINICA
2013年
6期
36-41
,共6页
李明想%柏素花%张玉刚%祝军%戴洪义
李明想%柏素花%張玉剛%祝軍%戴洪義
리명상%백소화%장옥강%축군%대홍의
苹果%甘露糖结合凝集素%基因克隆%表达分析
蘋果%甘露糖結閤凝集素%基因剋隆%錶達分析
평과%감로당결합응집소%기인극륭%표체분석
Malus domestica%Mannose-binding lectin(MBL)%Gene cloning%Gene expression
以苹果幼叶为试验材料,通过同源克隆和RT-PCR方法分离出B_LECTIN基因家族中的一个基因,命名为MdMBL。分析结果表明,克隆得到的MdMBL长度为1181 bp,包括20 bp的5′非编码区、81 bp的3′非编码区和一个长度为1080 bp编码359个氨基酸的开放阅读框,推测该蛋白质分子量为39.76 kDa,其等电点为8.72。在构建的MBL系统进化树中,MdMBL与葡萄的MBL的关系最近,其次为橹豆的MBL,与苹果的MBL2同源基因进化关系最远。相对荧光定量RT-PCR分析表明, MdMBL具有组织特异性,在茎中表达量最高,在花中表达量最低;低温胁迫促使MdMBL表达量升高,MdMBL在机械损伤处理下表达量先升高后降低;外源SA能够诱导MdMBL表达上调。
以蘋果幼葉為試驗材料,通過同源剋隆和RT-PCR方法分離齣B_LECTIN基因傢族中的一箇基因,命名為MdMBL。分析結果錶明,剋隆得到的MdMBL長度為1181 bp,包括20 bp的5′非編碼區、81 bp的3′非編碼區和一箇長度為1080 bp編碼359箇氨基痠的開放閱讀框,推測該蛋白質分子量為39.76 kDa,其等電點為8.72。在構建的MBL繫統進化樹中,MdMBL與葡萄的MBL的關繫最近,其次為艣豆的MBL,與蘋果的MBL2同源基因進化關繫最遠。相對熒光定量RT-PCR分析錶明, MdMBL具有組織特異性,在莖中錶達量最高,在花中錶達量最低;低溫脅迫促使MdMBL錶達量升高,MdMBL在機械損傷處理下錶達量先升高後降低;外源SA能夠誘導MdMBL錶達上調。
이평과유협위시험재료,통과동원극륭화RT-PCR방법분리출B_LECTIN기인가족중적일개기인,명명위MdMBL。분석결과표명,극륭득도적MdMBL장도위1181 bp,포괄20 bp적5′비편마구、81 bp적3′비편마구화일개장도위1080 bp편마359개안기산적개방열독광,추측해단백질분자량위39.76 kDa,기등전점위8.72。재구건적MBL계통진화수중,MdMBL여포도적MBL적관계최근,기차위로두적MBL,여평과적MBL2동원기인진화관계최원。상대형광정량RT-PCR분석표명, MdMBL구유조직특이성,재경중표체량최고,재화중표체량최저;저온협박촉사MdMBL표체량승고,MdMBL재궤계손상처리하표체량선승고후강저;외원SA능구유도MdMBL표체상조。
Mannose-binding Lectin(MBL),which belongs to B_LECTIN gene families,was isolated from the young leaf of apple ( Malus domestica) by homologous cloning technology ,named as MdMBL.The cDNA was 1 181 bp in length with an 5′non-coding region of 20 bp,an 3′non-coding region of 81 bp and an open reading frame ( ORF) of 1 080 bp encoding a protein of 359 amino acids ,and the estimated molecular weight and isoelectric point (pI)of the putative protein were 39.76 kDa and 8.72,respectively.The phylogenetic tree of MBL showed that Md-MBL was closest with grapes MBL ,followed by glycine max ,and the evolutionary relationships of homologous genes in apple MdMBL2 was farthest .Quantitative real-time PCR analysis shows that the gene is tissue-specific and ex-pressed highest in stems , the lowest in flowers;The expression of MdMBL was promoted increased with the cold stress and the mechanical damage stress;Exogenous SA could induce up-regulation of MdMBL expression .