Abstract
Brassica napus (oilseed rape) is an economically important oil crop worldwide. Sclerotinia stem rot (SSR) caused by Sclerotinia sclerotiorum is a threat to oilseed rape production. Because the flower petals play pivotal roles in the SSR disease cycle, it is useful to express the resistance-related genes specifically in flowers to hinder further infection with S. sclerotiorum. To screen flower-specific promoters, we first analyzed the transcriptome data from 12 different tissues of the B. napus line ZS11. In total, 249 flower-specific candidate genes with high expression in petals were identified, and the expression patterns of 30 candidate genes were verified by quantitative real-time transcription-PCR (qRT-PCR) analysis. Furthermore, two novel flower-specific promoters (FSP046 and FSP061 promoter) were identified, and the tissue specificity and continuous expression in petals were determined in transgenic Arabidopsis thaliana with fusing the promoters to β-glucuronidase (GUS)-reporter gene. GUS staining, transcript expression pattern, and GUS activity analysis indicated that FSP046 and FSP061 promoter were strictly flower-specific promoters, and FSP046 promoter had a stronger activity. The two promoters were further confirmed to be able to direct GUS expression in B. napus flowers using transient expression system. The transcriptome data and the flower-specific promoters screened in the present study will benefit fundamental research for improving the agronomic traits as well as disease and pest control in a tissue-specific manner.
Highlights
Brassica napus L. is an evolutionarily young allotetraploid species formed about 7500 years ago by natural hybridization between Brassica oleracea and Brassica rapa, followed by chromosome doubling [1,2]
The transcriptome data and the flower-specific promoters screened in the present study will benefit fundamental research for improving the agronomic traits as well as disease and pest control in a tissue-specific manner
FSP046 and FSP061 promoter can Drive GUS Expressing in Flowers of B. napus Based on the flower-specific expression in transgenic A. thaliana, we further investigated whether the GUBaSsegdenoencotuhled fbloewexepr-rsepsesceidficunedxeprreFsSsPio0n46inantdraFnSsPg0en61icpAro.mthoatleirandar,ivwinegfiunrtBh.enrapinuvseflsotiwgaetresd
Summary
Brassica napus L. is an evolutionarily young allotetraploid species formed about 7500 years ago by natural hybridization between Brassica oleracea and Brassica rapa, followed by chromosome doubling [1,2]. B. napus is an economically important oil crop all over the world. It provides vegetable oil for people, and provides high quality fodder for animals. Sclerotinia stem rot (SSR) is destructive to oilseed rape production. SSR is caused by Sclerotinia sclerotiorum (Lib.) de Bary, a cosmopolitan pathogen of many economically important crops. It infects more than 600 plant species, including important oil crops such as oilseed rape, soybean, and sunflower [3,4,5,6]. SSR deteriorates the quality of the seed, and significantly reduces the oil content [7]
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