紫花苜蓿MsNST的克隆及对木质素与纤维素合成的功能分析

蒋旭,崔会婷,王珍,张铁军,龙瑞才,杨青川,康俊梅

Cloning and Function Analysis of MsNST in Lignin and Cellulose Biosynthesis Pathway from Alfalfa

JIANG Xu,CUI HuiTing,WANG Zhen,ZHANG TieJun,LONG RuiCai,YANG QingChuan,KANG JunMei

表1 研究所用的引物序列

Table 1 Primer sequences used in this study

引物 Primer 核酸序列（5′-3′） Nucleotide sequence (5′-3′) 功能 Function 文献Reference MsNST-f TCAACTTTTTGGGTCCCTTGTG MsNST克隆 Cloning of MsNST MsNST-r TCACCACATGCTATCACCATTG actin-2s CAAAAGATGGCAGATGCTGAGGAT 内参基因 Reference gene Actin actin-2a CATGACACCAGTATGACGAGGTCG qNST-f TCATCTCAAAACCCTAGACAGCCC 实时定量PCR qRT-PCR qNST-r GTAATTTGCTTCATAATTCTCTTCCTTG Ntest-f GCACAATCCCACTATCCTTCG 转基因拟南芥阳性鉴定Identification of transgenic lines Ntest-r AGTTTTTTGATTTCACGGGTTGGGG W-NST-F CATTTGGAGAGAACACGGGGGACTCTAGAATGCCTGATAACATGAGTATAT Pbi121-MsNST超表达载体构建 Construction of Pbi121-MsNST overexpression vector W-NST-R AACATAAGGGACTGACCACCCGGGGATCCTCACCACATGCTTATCAC CATT Atact-f GCAACATACGACGAAATCAAGAA qRT-PCR内参基因 Actin gene for qRT-PCR [18] [19] [20] [21] Atact-r CGACACGAGAACTGTAACCCC AtPAL1-f ATGGAGATTAACGGGGCACAC 木质素与纤维素合成相关基因的表达分析 Expression analysis of lignin and cellulose biosynthesis related to genes AtPAL1-r GTACCGCCGAGAACACCGCC At4CL1-f GATTTGAGCTCGATAAGAGTGGTG At4CL1-r ATTTGCTAGTTTTGCCCTCA AtCESA4-f ATTCTGGGTGATTGGCGG AtCESA4-r AATAATGAGAGTTGTCGGAGGG AtCESA7-f TTCTTGCCTACTGTATCCTTCC AtCESA7-r GCTAACTCCGCTCCATCTCA AtCESA8-f CATCCCAACGCTATCAAACCTA AtCESA8-r CTGAGACACCTCCAATAACCCA MsCESA3-f TCGATGGGCTTTAGGTTCAG MsCESA3-r TGAGAAGAGGAATGGAAGTG MsCESA6-f CCCTCTTCATATCCATCGCAG MsCESA6-r CACCTCCAATCACCCAAAAC MsCESA7-f GATGAAGCAAGACAACCACTG MsCESA7-r CTGGGTTCATAAGTCTGTATCGG MsPAL-f ATGAGGTGAAGCGTATGGTG MsPAL-r CATCCCTAGCAGATTCAGACAG Ms4CL-f TTCACGTCCTTGCCTCATCA Ms4CL-r CCAAGTTTGTTGAGACCGGAGG MsCOMT-f AAAGTGATTGTGGCAGAATGCA MsCOMT-r TTTTGTGGCCAGGCTTGAA