The Journal of Horticultural Science & Biotechnology
Vol. 86 No: 6
Isolation and differential expression analysis of self-compatibility-related genes from mature pistils of 'Shatangju' mandarin (Citrus reticulata Blanco)
H.X. MIAO, Y.H. QIN, JAIME A. TEIXEIRA DA SILVA, Z.X. YE and G.B. HU
The objective of this research was to identify self-compatibility (SC)-related genes that are up-regulated in ‘Shatangju’ mandarin (Citrus reticulata Blanco). A suppression-subtractive hybridisation (SSH) cDNA library was constructed using self-compatible ‘Shatangju’ cDNA as the tester, and self-incompatible ‘Wuzishatangju’ mandarin cDNA as the driver.
Among the 949 positive clones obtained in the size range of 200 – 700 bp, 158 showed up-regulated expression in ‘Shatangju’ mandarin.
Several SC-related genes, such as those for the S1 self-incompatibility locus-linked pollen 3.15 protein (S1-protein), the ubiquitin-activating enzyme E1 (ubiquitin-E1), a zinc finger (C2H2 type) family protein (zinc-finger protein), the nuclear inhibitor of protein phosphatase-1 (nnpp-1), ubiquitin, molecular chaperones, and senescence-related proteins were identified.
Semi-quantitative reverse transcription-PCR (sqRT-PCR) and quantitative real-time PCR (qPCR) were used in this study to elucidate the differential expression patterns of five of these SC-related genes.
The results showed that expression levels of the S1-protein and of ubiquitin-E1 in ‘Shatangju’ ovaries and anthers were up-regulated by 60-fold and seven-fold, respectively, compared to the same tissues in ‘Wuzishatangju’ mandarin.
The expression levels of the zinc-finger protein and the ubiquitin protein in ‘Shatangju’ leaves and buds were higher than in leaves and buds of ‘Wuzishatangju’. Expression levels of the nnpp-1 gene in the filaments and anthers of ‘Shatangju’ were three-fold and four-fold higher, respectively, than in the same tissues of ‘Wuzishatangju’. These results suggest that the S1-protein and ubiquitin-E1 might play an important role in the self-compatible reaction of ‘Shatangju’ mandarin.
This information could provide new knowledge to understand SC, not only in citrus, but also in other fruit.
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