Genome-wide characterization and analysis of bHLH transcription factors related to anthocyanin biosynthesis in spine grapes (Vitis davidii)

Abstract

As one of the largest transcription factor family, basic helix-loop-helix (bHLH) transcription factor family plays an important role in plant metabolism, physiology and growth. Berry color is one of the important factors that determine grape quality. However, the bHLH transcription factor family’s function in anthocyanin synthesis of grape berry has not been studied systematically. We identified 115 bHLH transcription factors in grape genome and phylogenetic analysis indicated that bHLH family could be classified into 25 subfamilies. First, we screened six candidate genes by bioinformatics analysis and expression analysis. We found one of the candidate genes VdbHLH037 belonged to III (f) subfamily and interacted with genes related to anthocyanin synthesis through phylogenetic analysis and interaction network prediction. Therefore, we speculated that VdbHLH037 participated in the anthocyanin synthesis process. To confirm this, we transiently expressed VdbHLH037 in grape and Arabidopsis transformation. Compared with the control, transgenic materials can accumulate more anthocyanins. These results provide a good base to study the function of the VdbHLH family in anthocyanin synthesis of grape berry.

Genome-wide identification and characterization of PdbHLH transcription factors related to anthocyanin biosynthesis in colored-leaf poplar (Populus deltoids)

“…Genome-wide identification and characterization of bHLH family genes in many plants revealed that members of the III (f) subfamily have been proved to be involved in anthocyanin biosynthesis [ 28 , 29 , 54 ]. The FabHLH29 gene of white-flesh strawberry mutant, a member of the III (f) subfamily in bHLH TFs, has been reported to be involved in the anthocyanin biosynthesis [ 28 ]. In Freesia hybrida , two members of III (f) subfamily in bHLH TFs, FhGL3L and FhTT8L , contributed to the biosynthesis of flavonoids and anthocyanin [ 55 ].…”

“…Moreover, bHLH proteins of III (f) subfamily were involved in regulating the anthocyanin biosynthesis by interacting with MYB and WDR. In chrysanthemums, CmbHLH2 gene of III (f) subfamily regulates the anthocyanin biosynthesis by interacting with CmMYB6 to generate CmbHLH2 - CmMYB6 complex, which can bind to the promoter of CmDFR gene [ 28 ]. VvMYC1, the first bHLH TF described in the grapevine, can control anthocyanin and proanthocyanidin biosynthesis through interacting with MYB5a, MYB5b, MYBA1/A2 and MYBPA1 [ 56 ].…”

“…The genome-wide identification and characterization of bHLH gene family has been conducted in many plants [ 14 , 27 , 28 ]. There is 162 bHLH genes in Arabidopsis , 192 in tobacco, 159 in tomato, and 188 in apple, which can be divided into 15–26 subfamilies [ 28 ]. Among these subfamilies, members of the III subfamily have been proved to be involved in anthocyanin synthesis [ 29 ].…”

Systematic Analysis of bHLH Transcription Factors in Cassava Uncovers Their Roles in Postharvest Physiological Deterioration and Cyanogenic Glycosides Biosynthesis

“…The basic region, is a DNA-binding region that allows HLH proteins to bind to E-box (CANNTG) ( Massari and Murre, 2000 ). Basic helix-loop-helix transcription factors play crucial roles in plant growth and development including morphogenesis ( Gangappa and Kumar, 2017 ; Ballester et al, 2021 ), iron homeostasis ( Li et al, 2019 ), flower and fruit development ( Sun K. et al, 2019 ), stomatal initiation ( Raissig et al, 2016 ), root vascular cell proliferation ( Ohashi-Ito and Fukuda, 2016 ), grain yield ( Luo et al, 2013 ), secondary metabolites biosynthesis, such as anthocyanin ( Zhao R. et al, 2020 ; Li et al, 2021 ), also in the stress tolerance, for instance, drought ( Verma et al, 2020 ; Zhao Q. et al, 2020 ), salt ( Liu et al, 2021 ; Yu C. et al, 2021 ), cold ( Luo et al, 2020 ; Shen et al, 2021 ), heavy metal toxicity ( Sun W. et al, 2019 ), and osmotic stress ( Yang et al, 2016 ). Despite some bHLHs functions have been elaborated, most plant bHLHs functions are still unclear, especially in tuber crops.…”

Transcriptome Analysis Reveals Roles of Anthocyanin- and Jasmonic Acid-Biosynthetic Pathways in Rapeseed in Response to High Light Stress

“…More specifically, in Arabidopsis, three bHLH TF-encoding genes, GLABRA 3 ( GL3 ), ENHANCER OF GLABRA 3 ( EGL3 ), and TRANSPARENT TESTA 8 ( TT8 ), were demonstrated to participate in regulating anthocyanin biosynthesis [ 46 ]. Recently, the functions of bHLH s in regulating the anthocyanin-biosynthetic pathway have been investigated in various plant species, including apple, grape ( V. davidii ) [ 47 ], sheepgrass ( Leymus chinensis ) [ 48 ], kiwifruit ( Actinidia spp.) [ 49 ], and lotus ( Nelumbo nucifera ) [ 50 ].…”