Abstract:Multidrug and toxic compound extrusion transporters (MATE) are able to transport a wide variety of substrates such as metals, hormones and secondary metabolites, and they thus play an important role in growth and development in plant. In this study, the full-length coding sequence of MsMATE1 (NCBI GeneBank accessions: MN547958) and its promoter pMsMATE1 (MT505313) were isolated based on genome and transcriptome sequences of Medicago sativa under iron deficiency. The MsMATE1 is 1470 bp which encodes a putative protein of 489 amino acids. According to bioinformatics analysis, the MsMATE1 protein contains a typical MATE_like superfamily domain, which belongs to the eukaryotic subtype; MsMATE1 is a transmembrane protein, and the main component of its secondary structure is α helix. Phylogenetic analysis showed that MsMATE1 protein is closely related to MtMATE1 (XP013453190.1). The pMsMATE1 is 1598bp which were detected with multiple elements responding to hormone and abiotic stress. MsMATE1 was expressed in roots, shoots and leaves of alfalfa seedlings with the highest abundance in shoots. Under iron-deficient or iron-excess stresses, MsMATE1 was significantly up-regulated in different tissues of alfalfa seedlings, with the highest up-regulation in shoots. In MsMATE1 transgenic tobacco plants if compared with those of wild type plants, the activities of SOD, POD, and CAT, chlorophyll content and soluble protein content were significantly increased, while the significant reduction on content of MDA was observed under iron-deficient or iron-excess conditions, compared with those of wild type plants.The results showed that heterologous expression of MsMATE1 improved the tolerant against iron-deficient or iron-excess stresses in plants. MsMATE1 might be a candidate gene for improving tolerance to iron stress by genetic engineering. Collectively, this study laid a foundation for future deciphering the MsMATE1-involved molecular mechanism upon iron stresses.

Abstract:Floral development, directly related to the progeny reproduction and species continuation of plants, is an important stage in the lifespan of angiosperms. The MADS-box genes play a key role during the floral development. In this study, a new member of MADS-box family gene HaAGL11 was identified from the sunflower transcriptome datasets. Phylogenetic analysis showed that HaAGL11 gene was closely homologous with the AGL16 gene in Arabidopsis thaliana. Moreover, the expression pattern at different stages of flower development revealed that HaAGL11 gene was highly expressed at the late maturation stage of floret organs, flowering stage, and early embryonic development stage. The expression analysis in floret tissues indicated that HaAGL11 was highly expressing in the ovary of late maturation stage and flowering stage. Thus, this result suggested that HaAGL11 gene might be associated in regulating the development of ovary and fruit in the late stage of flower development and it could lay a foundation for preliminary exploration on the regulatory role of HaAGL11 gene in flower development and fruit formation of sunflower, beneficial for further investigating the molecular regulation mechanism of growth and development of sunflower, and providing some guidance data for the genetics and breeding of sunflower.

Abstract:CIPK (CBL-interacting protein kinase) is a serine / threonine protein kinase which plays animportant role in response to stress in plants. In this study, MsCIPK8 was obtained by using RT-PCR based ontranscriptional analysis of alfalfa in response to saline-alkaline stress. Sequence analysis of MsCIPK8 showed a1341 bp CDS encoding 446 amino acids with a relative molecular mass of 50.73 kDa and an isoelectric pointof 6.72. MsCIPK8 performed an N-terminal kinase domain and a C-terminal NAF/FISL domain as a CIPKSfamily protein. Bioinformatics analysis indicated that MsCIPK8 is a soluble protein with a high proportion ofrandom coil in secondary structure. Phylogenetic analysis showed that MsCIPK8 was most closely related toMtCIPK8 (Medicago truncatula).Four difference sites were found in the two protein sequence alignment, ofwhich 3 were in the conserved domain. MsCIPK8 had induced high expression in response to lowtemperature, drought, salt and salt-alkali. The expression of MsCIPK8 in roots and leaves had the highesttranscript abundance at 12 h and 3 h under low temperature stress respectively. Under saline stress, thehighest expression level of MsCIPK8 in roots was detected at 12 h. Under saline-alkaline stress, the expressionof MsCIPK8 in roots and leaves remained high after 12 h. Under drought stress, the expression of MsCIPK8 inroots and leaves reached the highest expression level at 12 h. These results indicated that the expression of MsCIPK8 might be related to the resistance of alfalfa to abiotic stresses such as drought, low temperature, salt and salt-alkali.

Abstract:Grain weight, as one of the three components, directly associates to the wheat yield production. Grain weight is recognized as a complex quantitative trait which is largely controlled by multiple functional genes/loci and which is also affected by genotype-environment interaction. The genes controlling the grain weight and its components are found to be distributed on eachchromosome of three wheat sub-genomes. Many molecular markers in wheat grain weight assisted selection have become applicable.The advances on the genetic characteristics, gene location, molecular mechanism and external factors affecting the grain weight formation have made with the contributions of national and international scholars. In this paper, the composing factors and influencing factors of wheat grain weight have been reviewed. The genetic localization, molecular cloning and allelic variation mining of the underlying genes are described, and the development of gene-based functional markers as well as the application in marker assisted breeding are summarized, as well as the future research prospects of wheat grain weight are proposed.

Abstract:Caffeoyl-CoA O-methyltransferase (CCoAOMT) is one of the key enzymes in lignin biosynthesis. By reversible transcription PCR (RT-PCR) technique, a CCoAOMT gene (DsCCoAOMT) was cloned from the bamboo shoot of Dendrocalamus sinicus. The full-length cDNA of DsCCoAOMT was 777 base pairs that encoded a protein of 238 amino acids with predicted molecular mass of 28.85kDa and a basic isoelectric point of 5.35. The deduced protein was rich in helix and coin. The phylogenetic analysis revealed the highest sequence homology of DsCCoAOMT in relative to CCoAOMT from the poaceous plants, followed by the monocotyledon of non-graminaceae and gymnosperms and then some dicotyledons. That suggested a differentiation of CCoAOMT between monocotyledons and dicotyledons before angiosperms evolution. According to the analysis of codon bias, the DsCCoAOMT had high bias toward the synonymous codons with G and C at third codon position. The results of codon usage frequency showed that heterologous expression of DsCCoAOMT might be applicable in the microorganism yeast expression system and in plants Nicotiana tabacum and Solanum lycopersicum. By real-time quantitative PCR, the DsCCoAOMT gene was highly expressed in the shoots, followed by the culms and the leaves at last. As the shoots developed, DsCCoAOMT is up-regulated at the higher level, suggesting that the gene might play an important regulation role in fast-growing D. sinicus. Thus, this study might be helpful for further understanding the function and molecular mechanism of the DsCCoAOMT gene.

Abstract:FTL (F-box Triple LRR ) protein belongs to the family of F-box proteins that play an important role in toleranceto low temperature. In this study, an MsFTL gene was isolated by candidate gene approach from alfalfa, and this gene wasdifferently expressed in leaf under cold stress. The full-length cDNA of MsFTL gene was 1422 bp, which putatively encoded for473 amino acids. By bioinformatic analysis, the MsFTL protein was found to carry an F-box domain and three LRR repeats inC-terminus. MsFTL showed homology closely to XP_003626345.1, a member of F-box/FBD/LRR-repeat protein in Medicagotruncatula, with 11 differential sites between the two proteins. The expression of MsFTL was induced by low temperature, salt,drought stress and ABA treatments. We generated the transformation construct and transformed the MsFTL into tobacco. Under-4°C, wild-type tobacco leaves showed remarkable wilting, while the transgenic plants presented slightly stressed phenotype.Transgenic plants accumulated higher content of soluble protein and soluble sugar than that of wild-type. The activity of SOD andCAT in transgenic plants was higher than those of wild type under low temperature (4°C) for 24 h. Moreover, the MDA content intransgenic plants was lower than that of wild type. Thus, these results suggested that overexpressing MsFTL might significantlyimprove tobacco plant tolerance to low temperature stress.

Abstract:This study reported the isolation of CiUPF0114 gene from Caragana intermedia, which contained an 876 bp open reading frame, encoding for a protein with 291 amino acids. The predicted isoelectric point and molecular weight of CiUPF0114 were 9.83 and 32.13 kD, respectively. The expression level of CiUPF0114 gene was significantly up-regulated under drought, dehydration or cold treatments. Furthermore by genome-wide screening, a total of 29 UPF0114 gene family members were identified from eight legumes, Arabidopsis and rice. The UPF0114 genes from were divided into subfamily A and subfamily B according to phylogenetic analysis. The A subfamily was further divided into two groups, A1 and A2. CiUPF0114, a gene from Caragana intermedia, belongs to the B subfamily. These results might lay the foundation for further study on the functions of CiUPF0114.

Abstract:Real-time fluorescence quantitative PCR is an effective method to quantify the transcriptional profile of target genes.Use of proximal gene as internal reference is essential when performing qRT-PCR experiments. The Actin (ACT) gene is highly conserved cross species and expressed stably and is often used as an internal control. In order to obtain the ACT gene of cauliflower,the ACT gene of cauliflower was cloned by RT-PCR method (GenBank ID: MG598643)1. The open reading frame (ORF) is 1,134 bp,encoding 377 amino acids with a predicted molecular weight of 41.77 kD and a hypothetical isoelectric point of 5.395. Wolf Psortanalysis indicated that BobActin protein was located in the cytoplasmic matrix, and Motif Scan analysis showed that BobActin proteinhad the conserved actin at position of 4-377 sites. BobActin shared 90% identity with the homologous proteins from genus Brassicacea,such as Brassica oleracea var. oleracea, Brassica rapa and Brassica napus. A pair of qRT-PCR primers was designed from theBobActin gene sequence, and this combination showed high specificity and amplification efficiency. qRT-PCR analysis indicated thatthe BobActin gene was stably expressed in tissues of cauliflower, including root, stem, flower ball, leaf, even under various stresstreatments (low temperature, high temperature, salt, drought and ABA). Thus, this gene might serve as an internal reference beingsuitable for gene expression in cauliflower.

Abstract:Glutathione S-transferases (GSTs) are superfamily of multifunctional enzymes which play important roles in stress tolerance and cellular detoxification. In this paper, a GST gene named BnGSTU1 (GenBank accession: MG941011) was isolated from ramie (Boehmeria nivea L.) by amplification of cDNA ends (RACE) based on the analysis of expression profiling of cadmium response genes in ramie. The open reading frame of BnGSTU1 is 636 bp, which encodes 211 amino acid residues. The predicted protein was 24.07 kD with isoelectic point of 5.29. Conserved domain search analysis revealed that BnGSTU1 protein contained a N-teminal domain and a C-teminal domain, as well as glutathione binding sites G and substrate binding pockets H, which showed typical characteristics of Tau class GSTs. Phylogenetic tree analysis showed that BnGSTU1 was closely to Tau class GST in Arabidopsis thaliana, Hevea brasiliensis and so on. Real-time quantitative PCR analysis indicated that BnGSTU1 expressed in root, stem and leaf of ramie, but the expression level of BnGSTU1 in leaf was significantly higher than that in roots. Furthermore, BnGSTU1 gene was significantly induced by cadmium stress, and the expression of BnGSTU1 increased along with Cd2+ concentration. Collectively, the results suggest that BnGSTU1 is a cadmium-responsive factor and may play potential roles in the plant adaption to cadmium stress.

Abstract:Improving the efficiency of nitrogen absorption in maize has important significance. In consideration of the characteristics of transportation of NO3- in CLC protein family, this study clones the CLC family genes ZmCLCa of maize by homology cloning method. Based on bioinformatics analysis, we find that the protein has a voltage-gated chlorine ion channel structure domain and subcellular localization result shows that the protein is located on the cell membrane. Under 200 mmol/L KNO3 processing conditions, the content of NO3- in transgenic arabidopsis of strain is obviously higher than that in wild arabidopsis. Therefore, ZmCLCa gene likely plays an important role in the process of nitrogen absorption of maize..