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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

Expression profiling of RuBisCo activase (TaRca1) in wheat during pollination and grain-filling stages under control (C) and heat stress (HS) conditions. (A) HD2329 (C and HS), (B) PBW621(C and HS), (C) Halna (C and HS), (D) HD2985 (C and HS); C −22 ± 3°, heat stress −42°C for 2 h; β-actin gene (accession no. AF282624) was used as endogenous control for normalizing the Ct value; Relative expression was calculated by Pfaffl method (Pfaffl et al., 2002); vertical bars indicate SE (n = 3).
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Figure 5: Expression profiling of RuBisCo activase (TaRca1) in wheat during pollination and grain-filling stages under control (C) and heat stress (HS) conditions. (A) HD2329 (C and HS), (B) PBW621(C and HS), (C) Halna (C and HS), (D) HD2985 (C and HS); C −22 ± 3°, heat stress −42°C for 2 h; β-actin gene (accession no. AF282624) was used as endogenous control for normalizing the Ct value; Relative expression was calculated by Pfaffl method (Pfaffl et al., 2002); vertical bars indicate SE (n = 3).

Mentions: HD2329 showed very high relative expression of TaRca1 in response to HS, as compared to control during pollination and grain-filling stages; maximum expression was observed during grain-filling in response to HS (Figure 5A). Similar pattern of expression of TaRca1 was observed in PBW621, HD2985, and Halna under control and HS-treated conditions (Figures 5B–D). Of all the cvs., relative expression of TaRca1 under HS was observed maximum in Halna and minimum in HD2329 during both the stages of growth. In general, abundance of TaRca1 transcript was observed during the grain-filling in all the four cultivars in response to HS.


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)

Expression profiling of RuBisCo activase (TaRca1) in wheat during pollination and grain-filling stages under control (C) and heat stress (HS) conditions. (A) HD2329 (C and HS), (B) PBW621(C and HS), (C) Halna (C and HS), (D) HD2985 (C and HS); C −22 ± 3°, heat stress −42°C for 2 h; β-actin gene (accession no. AF282624) was used as endogenous control for normalizing the Ct value; Relative expression was calculated by Pfaffl method (Pfaffl et al., 2002); vertical bars indicate SE (n = 3).
© Copyright Policy
Related In: Results  -  Collection

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

Figure 5: Expression profiling of RuBisCo activase (TaRca1) in wheat during pollination and grain-filling stages under control (C) and heat stress (HS) conditions. (A) HD2329 (C and HS), (B) PBW621(C and HS), (C) Halna (C and HS), (D) HD2985 (C and HS); C −22 ± 3°, heat stress −42°C for 2 h; β-actin gene (accession no. AF282624) was used as endogenous control for normalizing the Ct value; Relative expression was calculated by Pfaffl method (Pfaffl et al., 2002); vertical bars indicate SE (n = 3).
Mentions: HD2329 showed very high relative expression of TaRca1 in response to HS, as compared to control during pollination and grain-filling stages; maximum expression was observed during grain-filling in response to HS (Figure 5A). Similar pattern of expression of TaRca1 was observed in PBW621, HD2985, and Halna under control and HS-treated conditions (Figures 5B–D). Of all the cvs., relative expression of TaRca1 under HS was observed maximum in Halna and minimum in HD2329 during both the stages of growth. In general, abundance of TaRca1 transcript was observed during the grain-filling in all the four cultivars in response to HS.

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