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Analysis of Nicotiana tabacum PIN genes identifies NtPIN4 as a key regulator of axillary bud growth

Xiaodong Xie  Guangyong Qin  Ping Si  Zhaopeng Luo  Junping Gao  Xia Chen  Jianfeng Zhang  Pan Wei  Qingyou Xia  Fucheng Lin  Jun Yang  
【摘要】:The plant-specific PIN-FORMED(PIN) auxin efflux proteins have been well characterized in many plant species, where they are crucial in the regulation of auxin transport in various aspects of plant development. However, little is known about the exact roles of the PIN genes during plant development in Nicotiana species. This study investigated the PIN genes in tobacco(Nicotiana tabacum) and in two ancestral species(Nicotiana sylvestris and Nicotiana tomentosiformis). Genome-wide analysis of the N. tabacum genome identified 20 genes of the PIN family. An in-depth phylogenetic analysis of the PIN genes of N. tabacum, N. sylvestris and N. tomentosiformis was conducted.Nt PIN4 expression was strongly induced by the application of exogenous indole-3-acetic acid(IAA), but was downregulated by the application of ABA,a strigolactone analogue, and cytokinin, as well as by decapitation treatments,suggesting that the Nt PIN4 expression level is likely positively regulated by auxin. Expression analysis indicated that NtP IN4 was highly expressed in tobacco stems and shoots, which was further validated through analysis of the activity of the Nt PIN4 promoter. We used CRISPR-Cas9 technology to generate mutants for Nt PIN4 and observed that both T_0 and T_1 plants had a significantly increased axillary bud growth phenotype, as compared with the wild-type plants. Therefore, Nt PIN4 offers an opportunity for studying auxin-dependent branching processes.

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