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Identification of Putative RuBisCo Activase (TaRca1)-The Catalytic Chaperone Regulating Carbon Assimilatory Pathway in Wheat (Triticum aestivum) under the Heat Stress.

Kumar RR, Goswami S, Singh K, Dubey K, Singh S, Sharma R, Verma N, Kala YK, Rai GK, Grover M, Mishra DC, Singh B, Pathak H, Chinnusamy V, Rai A, Praveen S - Front Plant Sci (2016)

Bottom Line: HS causes decrease in the RuBisCo activity; maximum reduction was observed during pollination stage in thermosusceptible cvs. as validated through immunoblotting.We observed uniform carbon distribution in different tissues of thermotolerant cvs., as compared to thermosusceptible.There is, however, need to manipulate the thermal stability of TaRca1 enzyme through protein engineering for sustaining the photosynthetic rate under HS-a novel approach toward development of "climate-smart" crop.

View Article: PubMed Central - PubMed

Affiliation: Division of Biochemistry, Indian Agricultural Research Institute New Delhi, India.

ABSTRACT
RuBisCo activase (Rca) is a catalytic chaperone involved in modulating the activity of RuBisCo (key enzyme of photosynthetic pathway). Here, we identified eight novel transcripts from wheat through data mining predicted to be Rca and cloned a transcript of 1.4 kb from cv. HD2985, named as TaRca1 (GenBank acc. no. KC776912). Single copy number of TaRca1 was observed in wheat genome. Expression analysis in diverse wheat genotypes (HD2985, Halna, PBW621, and HD2329) showed very high relative expression of TaRca1 in Halna under control and HS-treated, as compared to other cultivars at different stages of growth. TaRca1 protein was predicted to be chloroplast-localized with numerous potential phosphorylation sites. Northern blot analysis showed maximum accumulation of TaRca1 transcript in thermotolerant cv. during mealy-ripe stage, as compared to thermosusceptible. Decrease in the photosynthetic parameters was observed in all the cultivars, except PBW621 in response to HS. We observed significant increase in the Rca activity in all the cultivars under HS at different stages of growth. HS causes decrease in the RuBisCo activity; maximum reduction was observed during pollination stage in thermosusceptible cvs. as validated through immunoblotting. We observed uniform carbon distribution in different tissues of thermotolerant cvs., as compared to thermosusceptible. Similarly, tolerance level of leaf was observed maximum in Halna having high Rca activity under HS. A positive correlation was observed between the transcript and activity of TaRca1 in HS-treated Halna. Similarly, TaRca1 enzyme showed positive correlation with the activity of RuBisCo. There is, however, need to manipulate the thermal stability of TaRca1 enzyme through protein engineering for sustaining the photosynthetic rate under HS-a novel approach toward development of "climate-smart" crop.

No MeSH data available.


Related in: MedlinePlus

Northern blot analysis of cloned RuBisCo activase (TaRca1) gene in contrasting wheat cvs. HD2985 and HD2329 at different stages of growth and development; [α-32P]-dCTP (300 μCi) were used for the probe labeling; 1.2% agarose gel was used for the electrophoretic separation of total RNA.
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Figure 3: Northern blot analysis of cloned RuBisCo activase (TaRca1) gene in contrasting wheat cvs. HD2985 and HD2329 at different stages of growth and development; [α-32P]-dCTP (300 μCi) were used for the probe labeling; 1.2% agarose gel was used for the electrophoretic separation of total RNA.

Mentions: The expression of cloned TaRca1 was further analyzed though northern blotting in the contrasting wheat cvs. HD2985 and HD2329 under control and HS-treated conditions. We observed significant increase in the expression of TaRca1 transcripts in HD2985 under HS, as compared with the control at all the stages studied; most prominent accumulation of transcript was observed during grain-filling (mealy-ripe) stage under HS (Figure 3). Similar pattern of expression was observed in cv. HD2329; the expression levels of TaRca1 under HS was, however, lower in HD2329 (thermosensitive) than in HD2985 (thermotolerant).


Identification of Putative RuBisCo Activase (TaRca1)-The Catalytic Chaperone Regulating Carbon Assimilatory Pathway in Wheat (Triticum aestivum) under the Heat Stress.

Kumar RR, Goswami S, Singh K, Dubey K, Singh S, Sharma R, Verma N, Kala YK, Rai GK, Grover M, Mishra DC, Singh B, Pathak H, Chinnusamy V, Rai A, Praveen S - Front Plant Sci (2016)

Northern blot analysis of cloned RuBisCo activase (TaRca1) gene in contrasting wheat cvs. HD2985 and HD2329 at different stages of growth and development; [α-32P]-dCTP (300 μCi) were used for the probe labeling; 1.2% agarose gel was used for the electrophoretic separation of total RNA.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4940427&req=5

Figure 3: Northern blot analysis of cloned RuBisCo activase (TaRca1) gene in contrasting wheat cvs. HD2985 and HD2329 at different stages of growth and development; [α-32P]-dCTP (300 μCi) were used for the probe labeling; 1.2% agarose gel was used for the electrophoretic separation of total RNA.
Mentions: The expression of cloned TaRca1 was further analyzed though northern blotting in the contrasting wheat cvs. HD2985 and HD2329 under control and HS-treated conditions. We observed significant increase in the expression of TaRca1 transcripts in HD2985 under HS, as compared with the control at all the stages studied; most prominent accumulation of transcript was observed during grain-filling (mealy-ripe) stage under HS (Figure 3). Similar pattern of expression was observed in cv. HD2329; the expression levels of TaRca1 under HS was, however, lower in HD2329 (thermosensitive) than in HD2985 (thermotolerant).

Bottom Line: HS causes decrease in the RuBisCo activity; maximum reduction was observed during pollination stage in thermosusceptible cvs. as validated through immunoblotting.We observed uniform carbon distribution in different tissues of thermotolerant cvs., as compared to thermosusceptible.There is, however, need to manipulate the thermal stability of TaRca1 enzyme through protein engineering for sustaining the photosynthetic rate under HS-a novel approach toward development of "climate-smart" crop.

View Article: PubMed Central - PubMed

Affiliation: Division of Biochemistry, Indian Agricultural Research Institute New Delhi, India.

ABSTRACT
RuBisCo activase (Rca) is a catalytic chaperone involved in modulating the activity of RuBisCo (key enzyme of photosynthetic pathway). Here, we identified eight novel transcripts from wheat through data mining predicted to be Rca and cloned a transcript of 1.4 kb from cv. HD2985, named as TaRca1 (GenBank acc. no. KC776912). Single copy number of TaRca1 was observed in wheat genome. Expression analysis in diverse wheat genotypes (HD2985, Halna, PBW621, and HD2329) showed very high relative expression of TaRca1 in Halna under control and HS-treated, as compared to other cultivars at different stages of growth. TaRca1 protein was predicted to be chloroplast-localized with numerous potential phosphorylation sites. Northern blot analysis showed maximum accumulation of TaRca1 transcript in thermotolerant cv. during mealy-ripe stage, as compared to thermosusceptible. Decrease in the photosynthetic parameters was observed in all the cultivars, except PBW621 in response to HS. We observed significant increase in the Rca activity in all the cultivars under HS at different stages of growth. HS causes decrease in the RuBisCo activity; maximum reduction was observed during pollination stage in thermosusceptible cvs. as validated through immunoblotting. We observed uniform carbon distribution in different tissues of thermotolerant cvs., as compared to thermosusceptible. Similarly, tolerance level of leaf was observed maximum in Halna having high Rca activity under HS. A positive correlation was observed between the transcript and activity of TaRca1 in HS-treated Halna. Similarly, TaRca1 enzyme showed positive correlation with the activity of RuBisCo. There is, however, need to manipulate the thermal stability of TaRca1 enzyme through protein engineering for sustaining the photosynthetic rate under HS-a novel approach toward development of "climate-smart" crop.

No MeSH data available.


Related in: MedlinePlus