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1. Identification of differentially expressed genes in 72 h styles from self-incompatible Citrus reticulata | |||
Miao Hongxia,Ye Zixing,Hu Guibing,Qin Yonghua | |||
Agronomy 18 July 2013 | |||
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Abstract:Self-incompatibility (SI) is one important factor that can result in seedless fruits of Citrus. Our previous study showed that 'Wuzishatangju' mandarin was gametophytic self-incompatibility (GSI) and 72 h was the crucial stage for the shift from a self-compatible to a self-incompatible state. However, it is not clear what genes are involved in the process. In this study, two SSH libraries were constructed to screen differentially expressed genes using 72 h styles after self-pollination of 'Wuzishatangju' and cross-pollination of 'Wuzishatangju' × 'Shatangju'. 106 and 97 differentially expressed cDNA clones were sequenced and identified from forward and reverse SSH libraries, respectively. Differentially expressed ESTs are possibly involved in the SI reaction of 'Wuzishatangju' through S-phase kinase-associated protein1 (SKP1)-like activity, autoinhibited Ca2+-ATPase 1, U-box domain protein, and serine/threonine kinase. Results from quantitative real-time PCR (qPCR) showed that SKP1-like and U-box were obviously up-regulated in ovaries before pollination of 'Wuzishatangju', and approximately 20-and 2-fold higher than that of 'Shatangju', respectively. The SKP1-like and U-box were obviously up-regulated in pistils at 4 d after self-pollination of 'Wuzishatangju', and approximately 20-and 800-fold higher than that the same stage after cross-pollination of 'Wuzishatangju' × 'Shatangju', respectively. The potential involvement of these genes in the SI reaction is discussed. | |||
TO cite this article:Miao Hongxia,Ye Zixing,Hu Guibing, et al. Identification of differentially expressed genes in 72 h styles from self-incompatible Citrus reticulata[J]. |
2. Extraction of high quality of RNA and construction of a suppression subtractive hybridization (SSH) library from Trifoliate Orange ( Poncirus trifoliata ) | |||
Jin-Zhi Zhang,Zhi-Min Li,Jia-Ling Yao,chun-Gen Hu | |||
Agronomy 21 January 2009 | |||
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Abstract:To gain a better understanding of gene expression in early flowering trifoliate orange mutant (precocious trifoliate orange, Poncirus trifoliata), suppressive subtractive hybridization (SSH) and bioinformatics were applied to identify differentially expressed genes in precocious trifoliate orange. However, isolation of high quality RNA from buds, internodal shoots, and mature leaf tissues of trifoliate orange is difficult because of its high levels of polyphenols, polysaccharides and other secondary metabolites. An modified extraction procedure gave satisfactory results, proteinase K and high concentrations of β- mercaptoethanol were used in the extraction buffer to improve RNA quality and yeild. In addition, proteinase K involved in the procedure avoided badly injury caused by diethyl pyrocarbonate (DEPC), too. RNA isolated using above method was of sufficient quality for construction of suppression subtraction hybridization (SSH) library, which allowed the isolation of several early flowering ESTs. | |||
TO cite this article:Jin-Zhi Zhang,Zhi-Min Li,Jia-Ling Yao, et al. Extraction of high quality of RNA and construction of a suppression subtractive hybridization (SSH) library from Trifoliate Orange ( Poncirus trifoliata )[OL].[21 January 2009] http://en.paper.edu.cn/en_releasepaper/content/28174 |
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