Clone and Expression Analysis of MePIL1 in Cassava

Abstract

Abstract: This work aims to clone MePIL1 gene from cassava and to reveal its functional roles in abiotic stresses（e.g.，shade），which will provide theoretical bases for optimizing the planting density of cassava. MePIL1 gene was cloned from leaf of cassava cultivar ‘Ku50’ by RT-PCR method，and the sequences of MePIL1 and its homology genes with other species were aligned and their phylogenetic tree was constructed. Subsequently，structural variations of MePIL1 were revealed between wild and cultivated cassava，and their expression patterns were investigated by quantitative RT-PCR. Results showed that MePIL1，which had a 1 578 bp open reading frame and encoded 525 amino acids and contained a HLH conserved domain，was cloned from cassava. Phylogenetic analysis showed that MePIL1 and homologous genes from Populus trichocarpa and Salix purpurea were clustered together，indicating that they had close genetic relationships. Analysis of gene structural variation revealed that the region of HLH domain was highly conserved，while most variations were derived from the differences between the wild and cultivated cassava species，and the distribution overall was relatively even. Expression pattern analysis showed that MePIL1 was highly expressed in leaf，and its expression was induced by osmotic stress and shade treatments. In conclusion，MePIL1 of cassava was successfully cloned and it was involved in the responses to shade and osmotic stresses at the transcriptional level in cassava.

Key words: cassava, MePIL1, shade stress, gene clone, expression analysis

CLC Number:

10.13560/j.cnki.biotech.bull.1985.2016.12.013

DING Ze-hong,TIE Wei-wei,FU Li-li,YAN Yan,XIA Zhi-qiang,WANG Wen-quan,HU Wei. Clone and Expression Analysis of MePIL1 in Cassava[J]. Biotechnology Bulletin, 2016, 32(12): 72-78.

References

[1] Mutsaers H, Ezumah H, Osiru D. Cassava-based intercropping：a review[J] . Field Crops Research, 1993, 34（3-4）：431-457.
[2] Johnston M, Onwueme I. Effect of shade on photosynthetic pigments in the tropical root crops：yam, taro, tannia, cassava and sweet potato[J] . Experimental Agriculture, 1998, 34（3）：301-312.
[3] Okoli PS, Wilson G. Response of cassava（Manihot esculenta Crantz）to shade under field conditions[J] . Field Crops Research, 1986, 14：349-359.
[4] Vandenbussche F, Pierik R, Millenaar FF, et al. Reaching out of the shade[J] . Current Opinion in Plant Biology, 2005, 8（5）：462-468.
[5] Salter MG, Franklin KA, Whitelam GC. Gating of the rapid shade-avoidance response by the circadian clock in plants[J] . Nature, 2003, 426（6967）：680-683.
[6] Zhang T, Maruhnich SA, Folta KM. Green light induces shade avoidance symptoms[J] . Plant Physiology, 2011, 157（3）：1528-1536.
[7] Nozue K, Tat AV, Devisetty UK, et al. Shade avoidance components and pathways in adult plants revealed by phenotypic profiling[J] . Plos Genet, 2015, 11（4）：e1004953.
[8] Li L, Zhang Q, Pedmale UV, et al. PIL1 participates in a negative feedback loop that regulates its own gene expression in response to shade[J] . Molecular Plant, 2014, 7（10）：1582-1585.
[9] Wang W, Feng B, Xiao J, et al. Cassava genome from a wild ancestor to cultivated varieties[J] . Nature Communications, 2014, 5：5110
[10] Bredeson JV, Lyons JB, Prochnik SE, et al. Sequencing wild and cultivated cassava and related species reveals extensive interspecific hybridization and genetic diversity[J] . Nature Biotechnology, 2016, 34：562-570.
[11] Wang H, Zhang Z, Li H, et al. CONSTANS-LIKE 7 regulates branching and shade avoidance response in Arabidopsis[J] . Journal of Experimental Botany, 2013, 64（4）：1017-1024.
[12] Tamura K, Peterson D, Peterson N, et al. MEGA5：molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods[J] . Molecular Biology and Evolution, 2011, 28（10）：2731-2739.
[13] Librado P, Rozas J. DnaSP v5：a software for comprehensive analysis of DNA polymorphism data[J] . Bioinformatics, 2009, 25（11）：1451-1452.
[14] Xu J, Duan X, Yang J, et al. Enhanced reactive oxygen species scavenging by overproduction of superoxide dismutase and catalase delays postharvest physiological deterioration of cassava storage roots[J] . Plant Physiology, 2013, 161（3）：1517-1528.
[15] 胡伟, 颜彦, 韦运谢, 等. 木薯MeASR基因克隆及表达分析[J] . 生物技术通报, 2015, 31（10）：125-130.
[16] Todaka D, Nakashima K, Maruyama K, et al. Rice phytochrome-interacting factor-like protein OsPIL1 functions as a key regulator of internode elongation and induces a morphological response to drought stress[J] . Proceedings of the National Academy of Sciences, 2012, 109（39）：15947-15952.
[17] Sakuraba Y, Jeong J, Kang MY, et al. Phytochrome-interacting transcription factors PIF4 and PIF5 induce leaf senescence in Arabidopsis[J] . Nature Communications, 2014, 5：4636.
[18] Hornitschek P, Kohnen MV, Lorrain S, et al. Phytochrome interacting factors 4 and 5 control seedling growth in changing light conditions by directly controlling auxin signaling[J] . The Plant Journal, 2012, 71（5）：699-711.