Cloning and Expression Analysis of miR156a-targeted Genes SPL6 and SPL9 in Camellia sinensis

LIU Yaqin; TIAN Kunhong; SUN Qilu; PAN Cheng; LI Yeyun; JIANG Jiayue; JIANG Changjun

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Journal of Tea Science ›› 2017, Vol. 37 ›› Issue (6) : 551-564.

Cloning and Expression Analysis of miR156a-targeted Genes SPL6 and SPL9 in Camellia sinensis

LIU Yaqin¹, TIAN Kunhong¹, SUN Qilu¹, PAN Cheng¹, LI Yeyun¹, JIANG Jiayue¹, JIANG Changjun^1,2,*

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Abstract

MicroRNA156a-SQUAMOSA promoter binding-like protein（miR156a-SPLs）play important roles in growth process, formation of leaves and response to stress in tea plant (Camellia sinensis). Based on transcriptome data, two cDNA CsSPL6 (2β318βbp) and CsSPL9 (1β954βbp) were cloned from tea plant. The nucleotide and amino acid sequences, biological information, phylogenetic tree and molecular expression models of the CsSPL6 and CsSPL9 were analyzed. Sequence analysis showed that the open reading frames of CsSPL6 and CsSPL9 are 1β668βbp and 1β116βbp, encoding 555 and 371 amino acids respectively. Both genes contain typical SBP domain and the recognition sites of microRNA156a. Quantitative real-time PCR analysis showed that the expression profile of Csn-miR156a was the highest in the buds, and the lowest in the 7th leaves, which was negatively correlated with CsSPL6 and CsSPL9. For different varieties(strains), the expression of Csn-miR156a was consistent with the growth of new shoots and the value of Photosynthesis index, but negatively correlated with the expression of CsSPL6 and CsSPL9, indicating. Csn-miR156a,CsSPL6 and CsSPL9 were involved in the growth of tea plant and can be used as markers to assess growth abilities among varieties. In addition, the expression of CsSPL9 was negatively correlatedwith Csn-miR156a under heat stress in four varieties (strains), which might be associated with the enhanced heat tolerance ability. The results showed that the expression of CsSPLs were negatively regulated by Csn-miR156a,which played an important role in the growth, development of tea plant and the stress resistance, providing a theoretical basis for understanding the growth and resistance mechanism of tea plant.

Key words

growth potential / high temperature, miR156a / SPL / Camellia sinensis

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LIU Yaqin, TIAN Kunhong, SUN Qilu, PAN Cheng, LI Yeyun, JIANG Jiayue, JIANG Changjun. Cloning and Expression Analysis of miR156a-targeted Genes SPL6 and SPL9 in Camellia sinensis[J]. Journal of Tea Science. 2017, 37(6): 551-564

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