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Multiple tissue-specific expression of rice seed-shattering gene SH4 regulated by its promoter pSH4.

Yan H, Ma L, Wang Z, Lin Z, Su J, Lu BR - Rice (N Y) (2015)

Bottom Line: Histochemical location and fluorescence analyses of GUS activity of transgenic plants indicated multiple tissue-specific expression of pSH4 in the seed-pedicel junction region of mature panicles (with highest level), stems, coleoptiles of germinated seeds, and scutella of mature seeds.The multiple tissue-specific expression pSH4 is categorized as a spatiotemporal promoter that drives the expression of the SH4 gene in different rice tissues, in addition to the seed-pedicel junction region.Our findings suggest that SH4 may have additional functions in the growth and development of rice, apart from its major role in seed shattering.

View Article: PubMed Central - PubMed

Affiliation: Ministry of Education Key Laboratory for Biodiversity and Ecological Engineering, Institute of Biodiversity Science, Fudan University, Songhu Road 2005, Shanghai, 200436 China.

ABSTRACT

Background: Rice seed shattering is an important domestication syndrome encoded by a gene named as SH4. The coding region of SH4 has been well studied regarding its function and roles in evolution. However, its promoter has not been identified, which limited our understanding of the detailed regulatory mechanisms of this gene. It is therefore critical to characterize the promoter and study its expression pattern.

Results: We analyzed the 5' upstream sequences of this gene and identified a ~2.6 kb fragment with typical promoter features, which was designated as pSH4. The promoter contained a number of cis-acting elements related to abscisic acid (ABA) and a CpG island that were characteristics of multiple tissue-specific expression. We isolated and ligated pSH4 to the β-glucuronidase (GUS) reporter gene, and transformed it into a japonica rice cultivar to determine the multiple expression pattern of SH4. Histochemical location and fluorescence analyses of GUS activity of transgenic plants indicated multiple tissue-specific expression of pSH4 in the seed-pedicel junction region of mature panicles (with highest level), stems, coleoptiles of germinated seeds, and scutella of mature seeds.

Conclusions: The multiple tissue-specific expression pSH4 is categorized as a spatiotemporal promoter that drives the expression of the SH4 gene in different rice tissues, in addition to the seed-pedicel junction region. Our findings suggest that SH4 may have additional functions in the growth and development of rice, apart from its major role in seed shattering.

No MeSH data available.


Related in: MedlinePlus

Structure of pSH4-GUS (A) and 35S-GUS (B) constructs for rice (Nipponbare) transformation. Tnos: terminator of nopaline synthetase; HYG(R): hygromycin-resistence gene; 35S: cauliflower mosaic virus (CaMV) 35S promoter; pSH4: SH4 promoter; XbaI and BglII indicate restriction sites; gusA: β-glucuronidase (GUS) gene; RB: right border; LB: left border.
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Fig6: Structure of pSH4-GUS (A) and 35S-GUS (B) constructs for rice (Nipponbare) transformation. Tnos: terminator of nopaline synthetase; HYG(R): hygromycin-resistence gene; 35S: cauliflower mosaic virus (CaMV) 35S promoter; pSH4: SH4 promoter; XbaI and BglII indicate restriction sites; gusA: β-glucuronidase (GUS) gene; RB: right border; LB: left border.

Mentions: The isolated fragment of potential promoter in the vector pMD18-pSH4 was inserted into the XbaI/BglII sites of vector pCAMBIA1301 to replace the CaMV35S promoter upstream from the GUS reporter gene to generate a recombinant vector, pSH4-GUS (Figure 6A). The inserted fragment was confirmed by PCR amplification with a primer pair (psh4aF: 5′-GCTGATCCGCTGGCCGTAGAAGTC-3′ and psh4aR: 5′-GCGCGTGAAGGGAGGGGGTTTA-3′) and sequenced with two primers (Pseq1: 5′-CCAGGCTTTACACTTTATGC-3′ and Pseq2: 5′-TTCACGGGTTGGGGTTTCTAC-3′). The pCAMBIA1301 vector without any inserts, where the GUS gene was still driven by a constitutive promoter CaMV35S, was used as the positive control for GUS analysis, and named as 35S-GUS (Figure 6B).Figure 6


Multiple tissue-specific expression of rice seed-shattering gene SH4 regulated by its promoter pSH4.

Yan H, Ma L, Wang Z, Lin Z, Su J, Lu BR - Rice (N Y) (2015)

Structure of pSH4-GUS (A) and 35S-GUS (B) constructs for rice (Nipponbare) transformation. Tnos: terminator of nopaline synthetase; HYG(R): hygromycin-resistence gene; 35S: cauliflower mosaic virus (CaMV) 35S promoter; pSH4: SH4 promoter; XbaI and BglII indicate restriction sites; gusA: β-glucuronidase (GUS) gene; RB: right border; LB: left border.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4384984&req=5

Fig6: Structure of pSH4-GUS (A) and 35S-GUS (B) constructs for rice (Nipponbare) transformation. Tnos: terminator of nopaline synthetase; HYG(R): hygromycin-resistence gene; 35S: cauliflower mosaic virus (CaMV) 35S promoter; pSH4: SH4 promoter; XbaI and BglII indicate restriction sites; gusA: β-glucuronidase (GUS) gene; RB: right border; LB: left border.
Mentions: The isolated fragment of potential promoter in the vector pMD18-pSH4 was inserted into the XbaI/BglII sites of vector pCAMBIA1301 to replace the CaMV35S promoter upstream from the GUS reporter gene to generate a recombinant vector, pSH4-GUS (Figure 6A). The inserted fragment was confirmed by PCR amplification with a primer pair (psh4aF: 5′-GCTGATCCGCTGGCCGTAGAAGTC-3′ and psh4aR: 5′-GCGCGTGAAGGGAGGGGGTTTA-3′) and sequenced with two primers (Pseq1: 5′-CCAGGCTTTACACTTTATGC-3′ and Pseq2: 5′-TTCACGGGTTGGGGTTTCTAC-3′). The pCAMBIA1301 vector without any inserts, where the GUS gene was still driven by a constitutive promoter CaMV35S, was used as the positive control for GUS analysis, and named as 35S-GUS (Figure 6B).Figure 6

Bottom Line: Histochemical location and fluorescence analyses of GUS activity of transgenic plants indicated multiple tissue-specific expression of pSH4 in the seed-pedicel junction region of mature panicles (with highest level), stems, coleoptiles of germinated seeds, and scutella of mature seeds.The multiple tissue-specific expression pSH4 is categorized as a spatiotemporal promoter that drives the expression of the SH4 gene in different rice tissues, in addition to the seed-pedicel junction region.Our findings suggest that SH4 may have additional functions in the growth and development of rice, apart from its major role in seed shattering.

View Article: PubMed Central - PubMed

Affiliation: Ministry of Education Key Laboratory for Biodiversity and Ecological Engineering, Institute of Biodiversity Science, Fudan University, Songhu Road 2005, Shanghai, 200436 China.

ABSTRACT

Background: Rice seed shattering is an important domestication syndrome encoded by a gene named as SH4. The coding region of SH4 has been well studied regarding its function and roles in evolution. However, its promoter has not been identified, which limited our understanding of the detailed regulatory mechanisms of this gene. It is therefore critical to characterize the promoter and study its expression pattern.

Results: We analyzed the 5' upstream sequences of this gene and identified a ~2.6 kb fragment with typical promoter features, which was designated as pSH4. The promoter contained a number of cis-acting elements related to abscisic acid (ABA) and a CpG island that were characteristics of multiple tissue-specific expression. We isolated and ligated pSH4 to the β-glucuronidase (GUS) reporter gene, and transformed it into a japonica rice cultivar to determine the multiple expression pattern of SH4. Histochemical location and fluorescence analyses of GUS activity of transgenic plants indicated multiple tissue-specific expression of pSH4 in the seed-pedicel junction region of mature panicles (with highest level), stems, coleoptiles of germinated seeds, and scutella of mature seeds.

Conclusions: The multiple tissue-specific expression pSH4 is categorized as a spatiotemporal promoter that drives the expression of the SH4 gene in different rice tissues, in addition to the seed-pedicel junction region. Our findings suggest that SH4 may have additional functions in the growth and development of rice, apart from its major role in seed shattering.

No MeSH data available.


Related in: MedlinePlus