Vol. 82 No: 1

Title:
Isolation and characterisation of the microspore-related gene BcMF4 in Chinese cabbage-pak-choi and its functional identification in Arabidopsis

Authors:
LECHENG LIU, YONGQIN WANG, TAO ZHANG, LI HUANG, XUN XIANG and JIASHU CAO

pp: 133-139

Abstract:
To provide information relevant to understanding the molecular mechanism of genic male-sterility in plants, differential expression of genes in flower buds at different developmental stages, and in other organs, between a sterile A line and a fertile B line of Chinese cabbage-pak-choi (Brassica rapa ssp. chinensis; syn. B. campestris ssp. chinensis) was analysed by cDNA-AFLP. Four cDNA fragments, including BcMF-A17T14, were obtained from flower buds of the fertile B line. In this paper, on the basis of BcMF-A17T14, a spliced full-length cDNA designated BcMF4 was generated by rapid amplification of cDNA ends (RACE) from flower buds of the fertile B line of Chinese cabbagepak- choi.The complete nucleotide sequence of the cDNA of BcMF4 was then amplified only from flower buds of the fertile B line. BcMF4 was expressed specifically in tetrads and uni-nucleate pollen of the fertile B line. The 1,102 bp DNA sequence of BcMF4 contains a single 95 bp intron and two exons. Its largest open reading frame (999 nt) encodes a polypeptide of 332 amino acids.The putative BcMF4 protein contains a signal peptide and displays five possible Nglycosylation sites, which are typical structures found in polygalacturonase inhibiting proteins (PGIPs). It also has eight Leucine-Rich Repeat (LRR) regions, and has 69 - 90% identity to PGIPs from other plants. Functional interruption of its ortholog in Arabidopsis thaliana, using an RNAi vector, resulted in aborted pollen grains and decreased pollen germination, and the abnormal pollen phenotype was stably inherited. This study suggests that the product of BcMF4 plays an important role in the development of microspores or pollen.

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