1.The responsive characteristics of phytochrome genes to photoperiod, abiotic stresses and identification of their key natural variation sites in foxtail millet (Setaria italica L.).
Xiaoping JIA ; Bo ZHANG ; Zhanxiang HE ; Jianfeng LI ; Xiaomei ZHANG ; Di GE ; Zhenshan WANG ; Luman SANG ; Zhiwei SONG
Chinese Journal of Biotechnology 2022;38(5):1929-1945
The responsive patterns of phytochrome gene family members to photoperiod and abiotic stresses were comparatively analyzed and the favorable natural variation sites of these genes were identified. This would help understand the mechanism of phytochrome gene family in photoperiod-regulated growth and development and abiotic stress response. In addition, it may facilitate the molecular marker assisted selection of key traits in foxtail millet. In this study, we used RT-PCR to clone three phytochrome genes SiPHYA, SiPHYB and SiPHYC from ultra-late maturity millet landrace variety 'Maosu'. After primary bioinformatics analysis, we studied the photoperiod control mode and the characteristics of these genes in responding to five abiotic stresses including polyethylene glycol (PEG)-simulated drought, natural drought, abscisic acid (ABA), high temperature and NaCl by fluorescence quantitative PCR. Finally, we detected the mutation sites of the three genes among 160 foxtail millet materials and performed haplotype analysis to determine the genes' functional effect. We found that the cloned cDNA sequences of gene SiPHYA, SiPHYB and SiPHYC were 3 981, 3 953 and 3 764 bp respectively, which contained complete coding regions. Gene SiPHYB and SiPHYC showed closer evolutionary relationship. Photoperiod regulated all of the three genes, but showed more profound effects on diurnal expression pattern of SiPHYB, SiPHYC than that of SiPHYA. Under short-day, when near heading, the expression levels of SiPHYA and SiPHYB were significantly lower than that under long-day, indicating their roles in suppressing heading of foxtail millet under long-day. SiPHYB and SiPHYC were responsive to PEG-simulated drought, natural drought, ABA and high temperature stresses together. SiPHYA and SiPHYB responded differently to salt stress, whereas SiPHYC did not respond to salt stress. Re-sequencing of 160 foxtail millet materials revealed that SiPHYB was highly conservative. Two missense mutations of SiPHYA, such as single nucleotide polymorphism (SNP) 7 034 522C→T and SNP7 036 657G→C, led to delaying heading and increasing plant height. One missense mutation of SiPHYC, such as SNP5 414 823G→T, led to shortening heading under short-day and delaying heading under long-day, as well as increasing plant height and panicle length regardless of photo-thermal conditions. Photoperiod showed different regulatory effects on SiPHYA, SiPHYB and SiPHYC. SiPHYB and SiPHYC jointly responded to various abiotic stresses except for the salt stress. Compared with the reference genotype, mutation genotypes of SiPHYA and SiPHYC delayed heading and increased plant height and panicle length.
Gene Expression Regulation, Plant
;
Photoperiod
;
Phytochrome/metabolism*
;
Plant Proteins/metabolism*
;
Setaria Plant/metabolism*
;
Stress, Physiological/genetics*
2.Phylogenetic and expression analysis of SEPALLATA-like gene in Brassica oleracea L. var. acephala.
Yuanping XIANG ; Yuntong HUANG ; Hongjun HE ; Qijiang XU
Chinese Journal of Biotechnology 2020;36(11):2398-2412
The E class MADS-box genes SEPALLATA (SEP)-like play critical roles in angiosperm reproductive growth, especially in floral organ differentiation. To analyze the sequence characteristics and spatio-temporal expression patterns of E-function MADS-box SEP-like genes during kale (Brassica oleracea L. var. acephala) flower development, BroaSEP1/2/3 (GenBank No. KC967957, KC967958, KC967960) homologues, three kale SEP MADS-box gene, were isolated from the kale variety 'Fourteen Line' using Rapid amplification of cDNA ends (RACE). Sequence and phylogenetic analysis indicated that these three SEP genes had a high degree of identity with SEP1, SEP2, SEP3 from Brassica oleracea var. oleracea, Brassica rapa, Raphanus sativus and Brassica napus, respectively. Alignment of the predicted amino acid sequences from these genes, along with previously published subfamily members, demonstrated that these genes comprise four regions of the typical MIKC-type MADS-box proteins: the MADS domain, intervening (I) domain and keratin-like (K) domain, and the C-terminal domain SEPⅠ and SEP Ⅱ motif. The longest open reading frame deduced from the cDNA sequences of BroaSEP1, BroaSEP2, and BroaSEP3 appeared to be 801 bp, 759 bp, 753 bp in length, respectively, which encoded proteins of 266, 252, and 250 amino acids respectively. Expression analyses using semi-quantitative RT-PCR and quantitative real-time PCR indicate that BroaSEP1/2/3 are specifically expressed in floral buds of kale during flower development process. The expression levels of the three genes are very different at different developmental stages, also in wild type, mutant flower with increased petals, and mutant flower with decreased petals. These different patterns of gene expression maybe cause the flowers to increase or decrease the petal number.
Brassica/metabolism*
;
Flowers/genetics*
;
Gene Expression Regulation, Plant
;
MADS Domain Proteins/metabolism*
;
Phylogeny
;
Plant Proteins/metabolism*
3.Genome-wide identification of BvHAK gene family in sugar beet (Beta vulgaris) and their expression analysis under salt treatments.
Xiaohan YANG ; Guoqiang WU ; Ming WEI ; Beichen WANG
Chinese Journal of Biotechnology 2022;38(10):3773-3789
High-affinity K+ transporter (HAK) is one of the most important K+ transporter families in plants and plays an important role in plant K+ uptake and transport. To explore the biological functions and gene expression patterns of the HAK gene family members in sugar beet (Beta vulgaris), physicochemical properties, the gene structure, chromosomal location, phylogenetic evolution, conserved motifs, three-dimensional structure, interaction network, cis-acting elements of promoter of BvHAKs were predicted by bioinformatic analysis, and their expression levels in different tissues of sugar beet under salt stress were analyzed by qRT-PCR. A total of 10 BvHAK genes were identified in the sugar beet genome. They contained 8-10 exons and 7-9 introns. The average number of amino acids was 778.30, the average molecular weight was 88.31 kDa, and the isoelectric point was 5.38-9.41. The BvHAK proteins contained 11-14 transmembrane regions. BvHAK4, -5, -7 and -13 were localized on plasma membrane, while others were localized on tonoplast. Phylogenetic analysis showed that HAK in higher plants can be divided into five clusters, namely cluster Ⅰ, Ⅱ, Ⅲ, Ⅳ, and Ⅴ, among which the members of cluster Ⅱ can be divided into three subclusters, including Ⅱa, Ⅱb, and Ⅱc. The BvHAK gene family members were distributed in cluster Ⅰ-Ⅳ with 1, 6, 1, and 2 members, respectively. The promoter of BvHAK gene family mainly contained stress responsive elements, hormone responsive elements, and growth and development responsive elements. The expression pattern of the BvHAK genes were further analyzed in different tissues of sugar beet upon salt treatment, and found that 50 and 100 mmol/L NaCl significantly induced the expression of the BvHAK genes in both shoots and roots. High salt (150 mmol/L) treatment clearly down-regulated their expression levels in shoots, but not in roots. These results suggested that the BvHAK gene family plays important roles in the response of sugar beet to salt stress.
Beta vulgaris/genetics*
;
Gene Expression Regulation, Plant
;
Phylogeny
;
Plant Roots
;
Sugars/metabolism*
;
Plant Proteins/metabolism*
4.Effect of straw apoplast protein on cellulase activity.
Di LU ; Hong-Zhang CHEN ; Run-Yu MA
Chinese Journal of Biotechnology 2006;22(2):257-262
We studied the alteration of the maize straw apoplast proteins in the process of preservation, and analyzed the effects of apoplast proteins on Penicillum expansum cellulase activities. The results show that: the extractable apoplast proteins are gradually decreased during the preservation of maize straw. Meanwhile, their synergistic effects on P. expensum cellulose are also attenuated. The apoplast proteins extracted from fresh maize straw possess endogenous EG activities, which is unstable and completely vanished after 6 months preservation. The apoplast proteins from the preserved straw exhibit significant synergistic effect on FPA, cotton lyase and beta-glucosidase. The maximal synergistic values are 95.32%, 102.06% and 96.6%, respectively. But interestingly, they inhibit the CMCase activity (max. 49.52%). Apoplast proteins show distinctive synergy with betaG and EG, but have no effect on CBH activity. After eliminating the effect of endogenous EG, the apoplast proteins from fresh maize straw have enhanced synergistic or inhibiting effects on FPA, Cotton lyase, betaG and CMCase than those extracted from the preserved straw. Based on our observation, the apoplast proteins play important roles in regulating the cellulase activities. The detailed analysis of the related mechanisms will greatly benefit the studies of the natural biomaterials hydrolysis.
Cellulase
;
metabolism
;
Penicillium
;
enzymology
;
Plant Proteins
;
metabolism
;
Plant Stems
;
metabolism
;
Zea mays
;
metabolism
5.Genome-wide identification of SUN gene family in Fragaria vesca and stresses-response analysis.
Yao YU ; Ziyao WANG ; Yiling XU ; Bojun MA ; Xifeng CHEN
Chinese Journal of Biotechnology 2023;39(2):724-740
SUN gene is a group of key genes regulating plant growth and development. Here, SUN gene families of strawberry were identified from the genome of the diploid Fragaria vesca, and their physicochemical properties, genes structure, evolution and genes expression were also analyzed. Our results showed that there were thirty-one FvSUN genes in F. vesca and the FvSUNs encoded proteins were classified into seven groups, and the members in the same group showed high similarity in gene structures and conservative motifs. The electronic subcellular localization of FvSUNs was mainly in the nucleus. Collinearity analysis showed that the members of FvSUN gene family were mainly expanded by segmental duplication in F. vesca, and Arabidopsis and F. vesca shared twenty-three pairs of orthologous SUN genes. According to the expression pattern in different tissues shown by the transcriptome data of F. vesca, the FvSUNs gene can be divided into three types: (1) expressed in nearly all tissues, (2) hardly expressed in any tissues, and (3) expressed in special tissues. The gene expression pattern of FvSUNs was further verified by quantitative real-time polymerase chain reaction (qRT-PCR). Additionally, the seedlings of F. vesca were treated by different abiotic stresses, and the expression level of 31 FvSUNs genes were assayed by qRT-PCR. The expression of most of the tested genes was induced by cold, high salt or drought stress. Our studies may facilitate revealing the biological function and molecular mechanism of SUN genes in strawberry.
Fragaria/metabolism*
;
Genes, Plant
;
Stress, Physiological/genetics*
;
Arabidopsis/genetics*
;
Plant Development
;
Gene Expression Regulation, Plant
;
Plant Proteins/metabolism*
6.Identification and expression analysis of MADS-box gene family in Docynia delavayi (Franch.) Schneid.
Xiwei WANG ; Can CHEN ; Dawei WANG
Chinese Journal of Biotechnology 2023;39(7):2897-2913
MADS-box gene family is a significant transcription factor family that plays a crucial role in regulating plant growth, development, signal transduction, and other processes. In order to study the characteristics of MADS-box gene family in Docynia delavayi (Franch.) Schneid. and its expression during different stages of seed germination, this study used seedlings at different stages of germination as materials and screened MADS-box transcription factors from the transcriptome database of D. delavayi using bioinformatics methods based on transcriptome sequencing. The physical and chemical properties, protein conservative motifs, phylogenetic evolution, and expression patterns of the MADS-box transcription factors were analyzed. Quantitative real-time PCR (qRT-PCR) was used to verify the expression of MADS-box gene family members during different stages of seed germination in D. delavayi. The results showed that 81 genes of MADS-box gene family were identified from the transcriptome data of D. delavayi, with the molecular weight distribution ranged of 6 211.34-173 512.77 Da and the theoretical isoelectric point ranged from 5.21 to 10.97. Phylogenetic analysis showed that the 81 genes could be divided into 15 subgroups, among which DdMADS27, DdMADS42, DdMADS45, DdMADS46, DdMADS53, DdMADS61, DdMADS76, DdMADS77 and DdMADS79 might be involved in the regulation of ovule development in D. delavayi. The combination of the transcriptome data and the qRT-PCR analysis results of D. delavayi seeds indicated that DdMADS25 and DdMADS42 might be involved in the regulation of seed development, and that DdMADS37 and DdMADS38 might have negative regulation effects on seed dormancy. Previous studies have reported that the MIKC* subgroup is mainly involved in regulating flower organ development. For the first time, we found that the transcription factors of the MIKC* subgroup exhibited a high expression level at the early stage of seed germination, so we speculated that the MIKC* subgroup played a regulatory role in the process of seed germination. To verify the accuracy of this speculation, we selected DdMADS60 and DdMADS75 from the MIKC* subgroup for qRT-PCR experiments, and the experimental results were consistent with the expression trend of transcriptome sequencing. This study provides a reference for further research on the biological function of D. delavayi MADS-box gene family from the perspective of molecular evolution.
MADS Domain Proteins/metabolism*
;
Phylogeny
;
Gene Expression Regulation, Plant
;
Genes, Plant
;
Transcription Factors/genetics*
;
Plant Proteins/metabolism*
;
Gene Expression Profiling
7.Research progress on effect of AP2/ERF transcription factors in regulating secondary metabolite biosynthesis.
Liang XIAO ; Rui-Bing CHEN ; Wu YU ; Lei ZHANG
China Journal of Chinese Materia Medica 2020;45(22):5412-5420
AP2/ERF transcription factor is a kind of transcription factors widely existing in plants, and contains at least a conserved AP2/ERF domain composed of about 60-70 amino acids. AP2/ERF transcription factors are widely involved in a variety of physiological processes in plants, including plant development, fruit ripening, flower development and other plant development processes, as well as such stress response processes as damage, pathogen defense, high-salt condition and drought. In recent years, secondary metabolic engineering that takes transcription factors as genetic manipulation targets has developed rapidly in improving the content of active ingredients and the quality of medicinal plants. This paper reviews the recent progress in the regulation of secondary metabolites biosynthesis with AP2/ERF transcription factors, and provides theoretical basis for the exploration of efficient regulatory targets, the regulation of secondary metabolites in medicinal plants, the targeted improvement of the content of active ingredients in traditional Chinese medicine, and the sustainable supply of high-quality traditional Chinese medicines.
Gene Expression Regulation, Plant
;
Phylogeny
;
Plant Proteins/metabolism*
;
Transcription Factor AP-2/metabolism*
;
Transcription Factors/metabolism*
8.Genome-wide analysis of LBD(lateral organ boundaries domain) gene family in Cannabis sativa of traditional Chinese medicine hemp seed.
Zhen WANG ; Mi YAO-LEI ; Xiang-Xiao MENG ; Hui-Hua WAN ; Ji AI-JIA ; Wei SUN ; Ma WEI
China Journal of Chinese Materia Medica 2020;45(22):5477-5486
LBD(lateral organ boundaries)transcription factors play an important role in the regulation of plant growth, development and secondary metabolism. In order to explore the function of LBD genes in cannabis, the Cannabis sativa genome and transcriptome were used to identify the C. sativa LBD gene family, and analyzed their expression patterns. Our results showed that the cannabis LBD contains 32 members, which were divided into two major categories, seven sub-families. Class Ⅰ was divided into 5 sub-families, named Class Ⅰ_a to Class Ⅰ_e, while Class Ⅱ was divided into 2 sub-families, including Class Ⅱ_a and Class Ⅱ_b. Analysis showed that the number of amino acids encoded LBDs was between 172 and 356, and the isoelectric point was between 4.92 and 9.43. The mole-cular weight of LBD was between 18 862.92 Da and 40 081.33 Da, and most members are located in the nucleus. Chromosome positioning of LBD showed that 32 members were unevenly distributed on 10 chromosomes of C. sativa LBD transcription factor domain, gene structure and motifs are relatively conservative, and the characteristics of different class members are similar. The upstream promoter region of the gene contains a variety of cis-acting elements related to plant hormones and environmental factors, C. sativa LBD genes have different expression patterns in the stems, leaves, and flowers of ZYS varieties(low tetrahydrocannabinol, high cannabidiol). The members of the LBD gene family are mainly expressed in the flowers and stems of ZYS varieties, while members expressed in the leaves very few; Class Ⅱ members CsLBD21 and CsLBD23 are expressed in flowers and stems, and CsLBD8 and CsLBD18 are expressed in flowers, stems and leaves. These genes may participate in the growth and development of cannabis and affect the biosynthesis of cannabinoids. This study laid the foundation for the subsequently functional research of the cannabis LBD gene family.
Cannabis/metabolism*
;
Gene Expression Regulation, Plant
;
Humans
;
Medicine, Chinese Traditional
;
Phylogeny
;
Plant Proteins/metabolism*
;
Seeds/metabolism*
9.Mining and identification of members of MYB transcription factor family in Lonicera macranthoides.
Juan ZENG ; Yu-Qing LONG ; Xue-Sen FU ; Ling WANG ; Zi-Xuan LIU ; Ri-Bao ZHOU ; Xiang-Dan LIU
China Journal of Chinese Materia Medica 2023;48(8):2103-2115
As a large family of transcription factors, the MYB family plays a vital role in regulating flower development. We studied the MYB family members in Lonicera macranthoides for the first time and identified three sequences of 1R-MYB, 47 sequences of R2R3-MYB, two sequences of 3R-MYB, and one sequence of 4R-MYB from the transcriptome data. Further, their physicochemical properties, conserved domains, phylogenetic relationship, protein structure, functional information, and expression were analyzed. The results show that the 53 MYB transcription factors had different conserved motifs, physicochemical properties, structures, and functions in wild type and 'Xianglei' cultivar of L. macranthoides, indicating their conservation and diversity in evolution. The transcript level of LmMYB was significantly different between the wild type and 'Xianglei' cultivar as well as between flowers and leaves, and some genes were specifically expressed. Forty-three out of 53 LmMYB sequences were expressed in both flowers and leaves, and 9 of the LmMYB members showed significantly different transcript levels between the wild type and 'Xianglei' cultivar, which were up-regulated in the wild type. The results provide a theoretical basis for further studying the specific functional mechanism of the MYB family.
Transcription Factors/metabolism*
;
Lonicera/metabolism*
;
Phylogeny
;
Plant Proteins/metabolism*
;
Gene Expression Regulation, Plant
10.Screening for UV-C irradiation-enhanced transcription factors that regulate the metabolism of phenolic compounds in tomato fruit.
Wenzhuo HAO ; Huanhuan ZHENG ; Changhong LIU ; Lei ZHENG
Chinese Journal of Biotechnology 2021;37(7):2483-2494
Solanum lycopersicum phenylalanine ammonia-lyase 5 (SlPAL5) gene regulates the metabolism of phenolic compounds. The study of transcription factors that regulate the expression of SlPAL5 gene is of great significance to elucidate the regulatory mechanism underlying the biosynthesis of phenolic compounds in tomato fruit induced by UV-C irradiation. Here, yeast one-hybrid library of tomato fruit was constructed, and the yeast one-hybrid technology was used to screen the transcription factors that regulate the expression of SlPAL5, the key gene related to the synthesis of phenolic compounds in tomato fruit. As a result, a transcription factor, SlERF7, was obtained and sequenced, followed by the blast homology analysis. Further experiments confirmed that SlERF7 interacted with the promoter of SlPAL5 gene. In addition, UV-C irradiation significantly increased the expression level of SlERF7. These results indicate that SlERF7, which is regulated by UV-C irradiation, might be involved in regulating the transcription of SlPAL5, which provided foundations for further studying the regulation mechanism of the biosynthesis of phenolic compounds in tomato fruit induced by UV-C irradiation.
Fruit
;
Gene Expression Regulation, Plant
;
Lycopersicon esculentum/metabolism*
;
Phenols
;
Plant Proteins/metabolism*
;
Transcription Factors/metabolism*