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.

References

1	WANG J Y， HU Z Z， ZHAO T M，et al.Genome-wide analysis of bHLH transcription factor and involvement in the infection by yellow leaf curl virus in tomato（Solanum lycopersicum）［J］.BMC Genomics，2015，16（1）：39.
2	GUO J R， SUN B X， HE H R，et al.Current understanding of bHLH transcription factors in plant abiotic stress tolerance［J］.International Journal of Molecular Sciences，2021，22（9）：4921.
3	MURRE C， BAIN G， VAN DIJK M A，et al.Structure and function of helix-loop-helix proteins［J］.Biochim Biophys Acta，1994 Jun 21；1218（2）：129-35.
4	王力伟，房永雨，刘红葵，等.bHLH转录因子的研究进展［J］.畜牧与饲料科学，2020，41（1）：23-27.
4	WANG L W， FANG Y Y， LIU H K，et al.Research progress of bHLH transcription factors［J］.Animal Husbandry and Feed Science，2020，41（1）：23-27.
5	XIE X B， LI S， ZHANG R F，et al.The bHLH transcription factor MdbHLH3 promotes anthocyanin accumulation and fruit colouration in response to low temperature in apples［J］.Plant，Cell & Environment，2012，35（11）：1884-1897.
6	ZHU J H， XIA D N， XU J，et al.Identification of the bHLH gene family in Dracaena cambodiana reveals candidate genes involved in flavonoid biosynthesis［J］.Industrial Crops & Products，2020，150：112407.
7	LIU W W， TAI H H， LI S S，et al. bHLH122 is important for drought and osmotic stress resistance in Arabidopsis and in the repression of ABA catabolism［J］.New Phytologist，2014， 201（4）：1192-1204.
8	SEO J S，JOO J， KIM M J，et al. OsbHLH148，a basic helix-loop-helix protein，interacts with OsJAZ proteins in a jasmonate signaling pathway leading to drought tolerance in rice［J］.The Plant Journal，2011，65（6）：907-921.
9	孙坤，丁雪洋，张辉，等.NBA1/MERIT40在3种沙棘中的适应性分化研究-兼论二代测序技术在杂种鉴定中的应用［J］.植物研究，2020，40（5）：648-658.
9	SUN K， DING X Y， ZHANG H，et al.Adaptive differentiation of NBA1/MERIT40 of three species of Hippophae L.-discussion on the application of next-generation sequencing technology in hybrid identification［J］.Bulletin of Botanical Research，2020，40（5）：648-658.
10	李雪丽.肋果沙棘和西藏沙棘自然杂交的转录组学研究［D］.兰州：西北师范大学，2021.
10	LI X L.Transcriptomics study on natural hybridization of Hippophae neurocarpa and H .thibetana［D］.Lanzhou：Northwest Normal University，2021.
11	张玉娜，孙坤，张辉，等.肋果沙棘（Hippophae neurocarpa S.W.Liu et T.N.He）自然种群的风媒传粉特征［J］.生态学报，2009，29（1）：508-514.
11	ZHANG Y N， SUN K， ZHANG H，et al.Characteristics of wind pollination in a natural population of Hippophae neurocarpa S.W.Liu et T.N.He（Elaeagnaceae）［J］.Acta Ecologica Sinica，2009，29（1）：508-514.
12	周璇，贾志鹏，王娟，等.肋果沙棘幼苗抗氧化酶系统及总黄酮对UV-B辐射增强的响应［J］.广西植物，2020，40（11）：1595-1601.
12	ZHOU X， JIA Z P， WANG J，et al.Response of antioxidant enzyme system and total flavonoid of woody plant Hippophae neurocarpa seedlings to enhanced UV-B radiation［J］.Guihaia，2020，40（11）：1595-1601.
13	温江波，孙坤，晏民生，等.青藏高原东缘西藏沙棘（Hippophae tibetana）果实性状变异［J］.植物研究，2010，30（2）：164-169.
13	WEN J B， SUN K， YAN M S，et al.Variation in fruit characteristics of Hippophae tibetana（Elaeagnaceae）in the eastern Qinghai-Tibet plateau，China［J］.Bulletin of Botanical Research，2010，30（2）：164-169.
14	赵雪艳，李思锋，柏国清，等.百合属5种植物的比较转录组分析［J］.植物研究，2021，41（4）：614-625.
14	ZHAO X Y， LI S F， BAI G Q，et al.Comparative transcriptome analysis of five species of Lilium ［J］.Bulletin of Botanical Research，2021，41（4）：614-625.
15	张莞晨，阮成江，李景滨，等.四种木本油料内参基因筛选及Actin基因时空表达分析［J］.分子植物育种，2018，16（14）：4576-4582.
15	ZHANG W C， RUAN C J， LI J B，et al.Screening of reference genes in four woody-oil trees and spatio-temporal expression analysis of actin gene［J］.Molecular Plant Breeding，2018，16（14）：4576-4582.
16	TARCZEWSKA A， GREB-MARKIEWICZ B.The significance of the intrinsically disordered regions for the functions of the bHLH transcription factors［J］.International Journal of Molecular Sciences，2019，20（21）：5306.
17	赵小明.家蚕丝腺特异表达的bHLH转录因子Bmsage和Bmdimm的功能研究［D］.重庆：西南大学，2014.
17	ZHAO X M.Function on BHLH transcription factors，Bmsage and Bmdimm，expressed specially in the silk gland of silkworm，Bombyx mori ［D］.Chongqing：Southwest University，2014.
18	PIRES N， DOLAN L.Origin and diversification of basic-helix-loop-helix proteins in plants［J］.Molecular Biology and Evolution，2010，27（4）：862-874.
19	HACHEZ C， OHASHI-ITO K， DONG J，et al.Differentiation of Arabidopsis guard cells：analysis of the networks incorporating the basic helix-loop-helix transcription factor，FAMA［J］.Plant Physiology，2011，155（3）：1458-1472.
20	DONG Y， WANG C P， HAN X，et al.A novel bHLH transcription factor PebHLH35 from Populus euphratica confers drought tolerance through regulating stomatal development，photosynthesis and growth in Arabidopsis［J］.Biochemical and Biophysical Research Communications，2014，450（1）：453-458.
21	ZHAO Q， FAN Z H， QIU L N，et al. MdbHLH130，an apple bHLH transcription factor，confers water stress resistance by regulating stomatal closure and ROS homeostasis in transgenic tobacco［J］.Frontiers in Plant Science，2020，11：543696.
22	王汉，乔枫.沙棘属逆境生理研究进展［J］.北方园艺，2021（9）：138-142.
22	WANG H， Qiao F.Research advances in stress physiology of Hippophae rhamnoides L.［J］.Northern Horticulture，2021（9）：138-142.
23	王昊.西藏沙棘的谱系地理学研究［D］.上海：复旦大学，2011.
23	WANG H.Phylogeography of Hippophae tibetana schlecht［D］.Shanghai：Fudan University，2011.
24	SUN X， WANG Y， SUI N.Transcriptional regulation of bHLH during plant response to stress［J］.Biochemical and Biophysical Research Communications，2018，503（2）：397-401.
25	WANG Y J， ZHANG Z G， HE X J，et al.A rice transcription factor OsbHLH1 is involved in cold stress response［J］.Theoretical and Applied Genetics，2003，107（8）：1402-1409.
26	ZHAO Q， XIANG X H， LIU D，et al.Tobacco transcription factor NtbHLH123 confers tolerance to cold stress by regulating the NtCBF pathway and reactive oxygen species homeostasis［J］.Frontiers in Plant Science，2018，9：381.
27	XU W R， ZHANG N B， JIAO Y T，et al.The grapevine basic helix-loop-helix（bHLH） transcription factor positively modulates CBF-pathway and confers tolerance to cold-stress in Arabidopsis ［J］.Molecular Biology Reports，2014，41（8）：5329-5342.
28	李小伟，张宏涛，张泽婧.西藏沙棘叶片黄酮含量与生态因子的相关性［J］.生态环境学报，2018，27（2）：239-245.
28	LI X W， ZHANG H T， ZHANG Z J.The relation between contents of flavonoids in leaves of Hippophae tibetana schlecht.and ecological factors［J］.Ecology and Environmental Sciences，2018，27（2）：239-245.
29	WANG X Y， JENSEN-TAUBMAN S M， KEEFE K M，et al.Achaete-scute complex homolog-1 promotes DNA repair in the lung carcinogenesis through matrix metalloproteinase-7 and O（6）-methylguanine-DNA methyltransferase［J］.PLoS One，2012，7（12）：e52832.
30	CARBONELL-BEJERANO P， DIAGO M P， MARTíNEZ-ABAIGAR J，et al.Solar ultraviolet radiation is necessary to enhance grapevine fruit ripening transcriptional and phenolic responses［J］.BMC Plant Biology，2014，14（1）：183.

[1]	Bo XU, Fu-Sun SHI, Li-Hua WANG, Zi-Song YANG. Effects of Altitude on Phenology and Morphological Characteristics of Fritillaria unibracteata [J]. Bulletin of Botanical Research, 2021, 41(5): 666-674.
[2]	SUN Kun, DING Xue-Yang, ZHANG Hui, LI Xue-Li, WANG Ying, WANG Juan, LIU Ben-Li. Adaptive Differentiation of NBA1/MERIT40 of Three Species of Hippophae L.——Discussion on the Application of Next-generation Sequencing Technology in Hybrid Identification [J]. Bulletin of Botanical Research, 2020, 40(5): 648-658.
[3]	MA Wen-Mei, WANG Yi-Feng, ZHAO Xia-Wei, LI Lei. Altitude Differences in Reproductive Characteristics and Resource Allocation of Saussurea wellbyi [J]. Bulletin of Botanical Research, 2019, 39(5): 707-715.
[4]	WANG Fei, TU Cai-Yun, CAO Xiu-Wen, LIU Jin-Qian, YANG Yong-Hong, ZHANG Tao, QI Hao. The Different Altitude Gradient Change Rules of the Main Shrub Community in Arid Valleys of the Bailongjiang River with Different Slope [J]. Bulletin of Botanical Research, 2018, 38(1): 26-36.
[5]	LI Jing-Hao, LI Hui, WEI Ya-Wei, ZHANG Song-Zhu, ZHU Wen-Xu, DENG Ji-Feng, SONG Yi-Xuan, ZHOU Yong-Bin. Water Use Efficiency of Pinus sylvestris, Pinus tabulaeformis, Quercus mongolica and Their Response Differences to Environmental Factors [J]. Bulletin of Botanical Research, 2016, 36(4): 581-587.
[6]	NING Hui-Xia;Nurbolt;LIU Ming;WANG Hui;KANG Xi-Liang;WANG Xiao-Jun. Biological Characteristics of Artificial Planting 1-Year-Old Saussurea involucrate* in Low Altitude Region [J]. Bulletin of Botanical Research, 2012, 32(6): 662-668.
[7]	SHI Hai-Yan;WANG Yi-Feng;WANG Jian-Hong. Studies on the Anatomical Structures of Leaves of Seven Saussurea* DC. Species on the Qinghai-Tibetan Plateau with Different Altitude [J]. Bulletin of Botanical Research, 2009, 29(3): 289-294.
[8]	MU Li-Qiang;MA Yao;YANG Guo-Ting;MA Da-Long. Anatomy structure and analysis of branches and leaves of Tilia amurensis in different altitude in Changbai mountain areas [J]. Bulletin of Botanical Research, 2006, 26(6): 658-662.
[9]	DUAN Xi-Hua, SUN Li-Fu, MA Shu-Rong, ZU Yuan-Gang. THE STUDIED OF BLADE OF ADENOPHORA POTANINII IN DIFFERENT ALTITUDE [J]. Bulletin of Botanical Research, 2003, 23(3): 334-336.
[10]	MA Shu-Rong, WU Suang-Xiu, WU Yu, YAN Xiu-Feng, ZU Yuan-gang. PATH COEFFICIENT ANALYSIS OF PHOTOSYNTHESIS AND TRANSPIRATION OF ADENOPHORA LOBOPHYLLA AND STOMATA BEHAVIOR [J]. Bulletin of Botanical Research, 2000, 20(4): 411-415.
[11]	Yan Ting-fen, Yan Xiu-feng, Zu Yuan-gang. A PRIMARYLY DISSCUSS ON THE ADAPTIVE MECHANISM AT DIFFERENT ALTITUDE LEVEL OF RHODIOLA SACHALINENSIS POPULATION [J]. Bulletin of Botanical Research, 1999, 19(2): 81-86.