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采用RT-PCR和RACE技术从‘香玲’核桃(Juglans regia L.‘Xiangling’)叶片中克隆了1个脱水素基因JrDHN(GenBank登录号KC503061.1),其全长1 056 bp,具有528 bp的开放阅读框,编码175个氨基酸组成的多肽,具有LEA家族成员的特征多肽序列,属于典型的Y2SK2型脱水素。以核桃18S基因为内参,对‘香玲’核桃JrDHN在4℃胁迫下的表达模式进行了初步的研究,结果表明低温诱导下叶片中JrDHN的表达增加,4℃处理4 h后达到最大值;自然越冬条件下,花芽中JrDHN表达量呈先上升后下降的趋势,在12月份表达量最高,推测JrDHN在核桃对低温胁迫响应中发挥重要功能。‘香玲’JrDHN在雌花中表达量高,其次是叶片和树皮,在花芽中最低。单核苷酸多态性分析JrDHN序列中有30个SNPs位点和11个Indels;单倍型分析显示12份核桃材料可分为10个单倍型,单倍型多样性为0.9697。
A dehydrin gene JrDHN (GenBank accession number KC503061.1) was cloned from leaves of Juglans regia L.’Xiangling ’by RT-PCR and RACE. The full-length cDNA was 1 056 bp in length with 528 bp open reading frame, which encodes a polypeptide of 175 amino acids and has the characteristic polypeptide sequence of LEA family member, belonging to the typical Y2SK2 type dehydratin. Using walnut 18S gene as an internal control, the expression pattern of JrDHN in ’Lingling’ walnut under 4 ℃ stress was studied. The results showed that the expression of JrDHN in leaves was increased under low temperature and reached the maximum at 4 ℃ for 4 h. Under natural winter conditions, the expression level of JrDHN in flower buds firstly increased and then decreased, and reached the highest level in December. It was speculated that JrDHN played an important role in the response of walnut to low temperature stress. ’Hong Ling’ JrDHN expressed high in female flowers, followed by leaves and bark, the lowest in flower buds. Single nucleotide polymorphism analysis JrDHN 30 SNPs sites and 11 Indels; haplotype analysis showed that 12 walnut materials can be divided into 10 haplotypes, haplotype diversity of 0.9697.