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Strength, Stability, and cis-Motifs of In silico Identified Phloem-Specific Promoters in Brassica juncea (L.)

Identifieur interne : 000050 ( Main/Exploration ); précédent : 000049; suivant : 000051

Strength, Stability, and cis-Motifs of In silico Identified Phloem-Specific Promoters in Brassica juncea (L.)

Auteurs : Murali Krishna Koramutla [Inde] ; Deepa Bhatt [Inde] ; Manisha Negi [Inde] ; Perumal Venkatachalam [Inde] ; Pradeep K. Jain [Inde] ; Ramcharan Bhattacharya [Inde]

Source :

RBID : PMC:4834444

Abstract

Aphids, a hemipteran group of insects pose a serious threat to many of the major crop species including Brassica oilseeds. Transgenic strategies for developing aphid-resistant plant types necessitate phloem-bound expression of the insecticidal genes. A few known phloem-specific promoters, in spite of tissue-specific activity fail to confer high level gene-expression. Here, we identified seven orthologues of phloem-specific promoters in B. juncea (Indian mustard), and experimentally validated their strength of expression in phloem exudates. Significant cis-motifs, globally occurring in phloem-specific promoters showed variable distribution frequencies in these putative phloem-specific promoters of B. juncea. In RT-qPCR based gene-expression study promoter of Glutamine synthetase 3A (GS3A) showed multifold higher activity compared to others, across the different growth stages of B. juncea plants. A statistical method employing four softwares was devised for rapidly analysing stability of the promoter-activities across the plant developmental stages. Different statistical softwares ranked these B. juncea promoters differently in terms of their stability in promoter-activity. Nevertheless, the consensus in output empirically suggested consistency in promoter-activity of the six B. juncea phloem- specific promoters including GS3A. The study identified suitable endogenous promoters for high level and consistent gene-expression in B. juncea phloem exudate. The study also demonstrated a rapid method of assessing species-specific strength and stability in expression of the endogenous promoters.


Url:
DOI: 10.3389/fpls.2016.00457
PubMed: 27148290
PubMed Central: 4834444


Affiliations:


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Le document en format XML

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<p>Aphids, a hemipteran group of insects pose a serious threat to many of the major crop species including
<italic>Brassica</italic>
oilseeds. Transgenic strategies for developing aphid-resistant plant types necessitate phloem-bound expression of the insecticidal genes. A few known phloem-specific promoters, in spite of tissue-specific activity fail to confer high level gene-expression. Here, we identified seven orthologues of phloem-specific promoters in
<italic>B. juncea</italic>
(Indian mustard), and experimentally validated their strength of expression in phloem exudates. Significant
<italic>cis</italic>
-motifs, globally occurring in phloem-specific promoters showed variable distribution frequencies in these putative phloem-specific promoters of
<italic>B. juncea</italic>
. In RT-qPCR based gene-expression study promoter of
<italic>Glutamine synthetase 3A (GS3A)</italic>
showed multifold higher activity compared to others, across the different growth stages of
<italic>B. juncea</italic>
plants. A statistical method employing four softwares was devised for rapidly analysing stability of the promoter-activities across the plant developmental stages. Different statistical softwares ranked these
<italic>B. juncea</italic>
promoters differently in terms of their stability in promoter-activity. Nevertheless, the consensus in output empirically suggested consistency in promoter-activity of the six
<italic>B. juncea</italic>
phloem- specific promoters including GS3A. The study identified suitable endogenous promoters for high level and consistent gene-expression in
<italic>B. juncea</italic>
phloem exudate. The study also demonstrated a rapid method of assessing species-specific strength and stability in expression of the endogenous promoters.</p>
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