Isolation and Analysis of Differentially Expressed Genes

between Male Fertile and Male Sterile Flower buds

OF MARIGOLD (TAGETES ERECTA L.)

 

Zhiqiang Hou^1,2, Daocheng Tang² AND Nan Tang^{1, 2*}

 

¹State Key Laboratory of Plateau Ecology and Agriculture, College of Agriculture and Animal Husbandry,

Qinghai University, 810016 Xining, China

²Key Laboratory of Qinghai Province for Landscape Plants Research, Plateau Flower Research Center,

Qinghai University, 810016 Xining, China

*Corresponding author’s email: natasha_tn@hotmail.com; telephone: (86)09715362782

 

Abstract

 

Male sterility is an important approach in utilization of heterosis in marigold (Tagetes erecta L.). Study on the mechanism of male sterility is very important, especially in mining of fertility-related genes. Three suppression subtractive hybridization (SSH) cDNA libraries were constructed between male fertile and male sterile flower buds of marigold. Out of 1920 clones, five hundred and six positive clones were verified by dot-blot hybridization. Two hundred and eighty-six non-redundant ESTs were obtained of which, one hundred and ninety-two ESTs corresponding to proteins with known functions. Through GO function annotation, fifteen candidate genes that may have a function in male sterility were identified. These genes involved in hormone pathways and cell cycles as well as encoded transcription factors and protein kinases. Further more, five of them were verified by quantitative real-time PCR, they were CDKB2;1 functioned in cell division, AMS involved in anther wall tapetum development, LAP3 played a role in pollen exine formation, ACOS5 and CYP703A2 involved in sporopollenin biosynthetic process. This is the first study that constructing cDNA libraries containing differentially expressed gene pools associate with male fertility using SSH strategy, and provides a first step to understand the mechanism of male reproductive development in marigold.

 

Key words: Tagetes erecta L., Male sterility, Suppression subtractive hybridization, Differentially expressed genes, Quantitative real-time PCR.