Study on Adaptive Differentiation of Transcription Factor bHLH94 Gene to Altitude in Hippophae neurocarpa and H.tibetana

doi:10.7525/j.issn.1673-5102.2022.06.007

Abstract

Abstract:

The bHLH proteins are the second large family of transcription factors， which play important roles in plant growth development， physiological metabolism and stress responses. In this study， Hippophae neurocarpa and H.tibetana were used as materials， the positive selection of the transcription factor bHLH94 genewas screened out using transcriptome sequencing. Based on the analysis of gene sequence and expression of HnbHLH94 and HtbHLH94， the response mechanism of bHLH94 genein H.neurocarpa and H.tibetana adapted to altitude was studied. The HnbHLH94 and HtbHLH94 gene encoded 338 and 335 amino acids respectively. Sanger sequencing method verified the correctness of sequences， however there were 10 non-synonymous mutation sites outside the DNA binding domain of the two genes， which was speculated to be related to the adaptive evolution of the gene. As the altitude increased， the HnbHLH94 gene expression decreased， while the HtbHLH94 gene expression increased based on qRT-PCR technology. It suggested that gene might provide a molecular basis for adaptation to altitude， drought， freezing and radiation respectively， and the sequence structure and expression of HnbHLH94 and HtbHLH94 genes responded to the habitat conditions of elevation.

Key words: bHLH94 transcription factor, Hippophae neurocarpa, H.tibetana, altitude, adaptive differentiation

CLC Number:

S793.6

Ting QIAN, Fan ZHAO, Yujie ZHANG, Xueli LI, Kun SUN, Hui ZHANG. Study on Adaptive Differentiation of Transcription Factor bHLH94 Gene to Altitude in Hippophae neurocarpa and H.tibetana[J]. Bulletin of Botanical Research, 2022, 42(6): 976-985.

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物种 Species	采集地 Location	样品编号 Code. No	海拔 Altitude /m
肋果沙棘 H. neurocarpa	青海省果洛州达日桥头 Dari Bridge Head，Goluo Prefecture，Qinghai Province	N_H	3 895
	青海省野牛沟乡大浪村 Dalang Village，Ye niugou Township，Qinghai Province	N_M	3 533
	青海省祁连县八宝镇冰沟村 Binggou Village，Babao Town，Qilian County，Qinghai Province	N_ML	3 352
	青海省祁连县日旭村 Rixu Village，Qilian County，Qinghai Province	N_L	3 188
西藏沙棘 H. tibetana	青海省果洛州达日县东日寺 Dongri Temple，Dari County，Guoluo Prefecture，Qinghai Province	T_H	4 066
	青海省野牛沟乡大浪村 Dalang Village，Yeniugou Township，Qinghai Province	T_M	3 533
	青海省祁连县日旭村 Rixu Village，Qilian County，Qinghai Province	T_L	3 188

物种 Species	NCBI物种编号 Number of species in NCBI	数据来源 Data sources	蛋白序列版本 Version of protein sequence
Amborella trichopoda	txid13333	https：//phytozome.jgj.doe.gov/pz/portal.html	v1.0
Arabidopsis thaliana	txid3702	https：//phytozome.jgj.doe.gov/pz/portal.html	TAIR10
Zea mays	txid4577	https：//phytozome.jgj.doe.gov/pz/portal.html	v1.1
Helianthus annuus	txid4232	https：//phytozome.jgj.doe.gov/pz/portal.html	r1.2
Solanum lycopersicum	txid4081	https：//phytozome.jgj.doe.gov/pz/portal.html	ITAG2.4
Oryza sativa	txid4530	https：//phytozome.jgj.doe.gov/pz/portal.html	v7.0
Physcomitrium patens	txid3128	https：//phytozome.jgj.doe.gov/pz/portal.html	v3.3
Selaginella moellendorffii	txid88036	https：//phytozome.jgj.doe.gov/pz/portal.html	v1.0
Picea abies	txid3329	https：//congenie.org/	v1.0
Ziziphus jujuba	txid26968	https：//www.ncbi.nlm.nih.gov/	1.1
Trema orientale	txid63057	https：//www.ncbi.nlm.nih.gov/	asm01
Parasponia andersonii	txid3476	https：//www.ncbi.nlm.nih.gov/	asm01
Chlamydomonas reinhardtii	txid3055	https：//www.ncbi.nlm.nih.gov/	v3.0
Hippophae neurocarpa	txid193523	本研究提供 Provied from this research
Hippophae tibetana	txid193519	本研究提供 Provied from this research

样品 Sample	200~500 bp	500~1 kbp	1 k~2k bp	>2 kbp	合计 Total
N_L	191 173	29 975	12 141	6 310	239 599
N_ML	158 804	26 688	11 272	6 342	203 106
N_M	183 449	28 847	11 833	6 218	230 347
N_H	151 730	24 374	10 996	5 853	192 953
T_L	155 163	28 033	12 361	6 689	202 246
T_M	230 291	41 293	17 601	8 177	297 362
T_H	151 032	26 931	11 340	6 269	195 572

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