摘要
利用徒手切片法、pH差计法及实时荧光定量RT-PCR技术,观察和测定紫心大白菜与同源非紫心大白菜[Brassica campestris L.ssp.pekinensis(Lour)Olsson]花青素在叶片中的分布和总含量变化,比较叶片中花青素合成途径关键酶基因及其转录调控因子的表达特点。结果表明,紫心大白菜花青素不均匀分布于叶片中,尤以叶表皮细胞及临近表皮的叶肉细胞内最为丰富,并且内层球叶含量高于外层球叶。紫心大白菜心叶中花青素含量最高为0.79mg·g-1FW,对照09S17混合样的叶片中花青素总含量为0.01mg·g-1FW。荧光定量PCR分析表明,花青素生物合成途径上游关键酶基因CHS、CHI、F3H、F3′H及下游修饰基因UFGT、转运酶基因GST与转录因子MYB0在紫色大白菜心叶中上调表达,下游关键酶基因DFR、ANS、LDOX与转录因子MYB2、MYB4、MYB12和MYB111的表达在整个紫心大白菜中大幅上调,推测这可能是紫心大白菜花青素积累的重要原因,其中MYB2和MYB4可能起主要作用。
To investigate the mechanism of anthocyanin production in our new purple-heading Chinese cabbage[Brassica campestris L. ssp. pekinensis（Lour）Olsson],the expression of structural genes and regulatory genes involved in anthocyanin biosynthesis was examined between the purple line （11S96）and its non-purple parent（09S17）by the real-time fluorescent quantitative RT-PCR,as well as their leaves cross-section were observed and the total anthocyanin content was measured. Leaf cross-section showed that the anthocyanin distribution is non-uniform in the purple heading leaf,specially enriched in the epidermis and its adjacent mesophyll cells. Comparison with the negligible amount of anthocyanin in the non-purple parent,the anthocyanin in purple line is up to 0.79 mg · g^-1 FW in purple inner heading leaves. Q-PCR analysis indicated that the EBGs（Early Biosynthesis Genes）CHS,CHI,F3H,F3′H,LBGs（Late Biosynthesis Genes）UFGT and GST,as well as the regulatory gene MYB0 were up-regulated in the inner leaves of purple heading;the other three LBGs,DFR,ANS,LDOX and the regulatory genes MYB2,MYB4,MYB12 and MYB111 were high up-regulated in all the leaves of the purple heading,which means that there is probably close relationship between these genes and the anthocyanin accumulation in this purple-heading Chinese cabbage,and MYB2 and MYB4 may play the dominant role in it.
出处
《园艺学报》
CAS
CSCD
北大核心
2012年第11期2159-2167,共9页
Acta Horticulturae Sinica
基金
国家‘863’计划项目(2012AA100205-5)
国家自然科学基金项目(31171965)
陕西省蔬菜产业体系项目
关键词
大白菜
紫心
花青素
分布
基因表达分析
Chinese cabbage, purple-heading, anthocyanin, distribution, gene expression analysis
