WRKY proteins are members of a family of transcription factors in higher plants that function in plant responses to various physiological processes. We identified 120 candidate WRKY genes from Gossypium raimondii with corresponding expressed sequence tags in at least one of four cotton species, Gossypium hirsutum, Gossypium barbadense, Gossypium arboreum, and G. raimondii. These WRKY members were anchored on 13 chromosomes in G. raimondii with uneven distribution. Phylogenetic analysis showed that WRKY candidate genes can be classified into three groups, with 20 members in group I, 88 in group II, and 12 in group III. The 88 genes in group II were further classified into five subgroups, groups IIa-e, containing 7, 16, 37, 15, and 13 members, respectively. We characterized diversity in amino acid residues in the WRKY domain and/or other zinc finger motif regions in the WRKY proteins. The expression patterns of WRKY genes revealed their important roles in diverse functions in cotton developmental stages of vegetative and reproductive growth and stress response. Structural and expression analyses show that WRKY proteins are a class of important regulators of growth and development and play key roles in response to stresses in cotton.
Plant microRNAs (miRNAs) play important roles in biological processes such as development and stress responses. Although the diverse functions of miRNAs in model organisms have been well studied, their function in wild rice is poorly understood. In this study, high-throughput small RNA sequencing was performed to characterize tissue-specific transcriptomes in Oryza longistaminata. A total of 603 miRNAs, 380 known rice miRNAs, 72 conserved plant miRNAs, and 151 predicted novel miRNAs were identified as being expressed in aerial shoots and rhizomes. Additionally, 99 and 79 miRNAs were expressed exclusively or differentially, respectively, in the two tissues, and 144 potential targets were predicted for the differentially expressed miRNAs in the rhizomes. Functional annotation of these targets suggested that transcription factors, including squamosa promoter binding proteins and auxin response factors, function in rhizome growth and development. The expression levels of several miRNAs and target genes in the rhizomes were quantified by RT-PCR, and the results indicated the existence of complex regulatory mechanisms between the miRNAs and their targets. Eight target cleavage sites were verified by RNA ligase-mediated rapid 5′ end amplification. These results provide valuable information on the composition, expression and function of miRNAs in O. longistaminata, and will aid in understanding the molecular mechanisms of rhizome development.
Groundnut (Arachis hypogaea L.) is widely grown and consumed around the world and is considered to have originated from a single hybridization event between two wild diploids. The utilization of wild germplasm in breeding programs has been restricted by reproductive barriers between wild and cultivated species and technical difficulties in making large numbers of crosses. Efforts to overcome these hurdles have resulted in the development of synthetic amphidiploids, namely ISATGR 278-18 (Arachis duranesis × Arachis batizocoi) and ISATGR 5B (Arachis magna × A. batizocoi), which possess several desirable traits, including resistance to foliar diseases that generally cause huge yield losses annually in groundnut growing areas of Asia, America, and Africa. With an objective to improve foliar disease resistance, the primary gene pool was diversified by introgressing foliar disease resistance in five cultivated genotypes (ICGV 91114, ICGS 76, ICGV 91278, JL 24, and DH 86) from synthetic amphidiploids using a backcross breeding approach. Several introgression lines with resistance to two foliar diseases (rust and late leaf spot) were identified with levels of resistance equal to the donors. These backcross derived lines have shown a wide range of variation for several morphological and agronomic traits. These lines, after further evaluation and selection, can serve as donors in future breeding programs aimed at developing improved cultivars with desirable agronomic traits, high resilience to biotic/abiotic stresses and a broadened genetic base.
The transcription factor dehydration-responsive element binding protein (DREB) is able to improve tolerance to abiotic stress in plants by regulating the expression of downstream genes involved in environmental stress resistance. The objectives of this study were to evaluate the salt tolerance of GmDREB1 transgenic wheat (Triticum aestivum L.) and to evaluate its physiological and protein responses to salt stress. Compared with the wild type, the transgenic lines overexpressing GmDREB1 showed longer coleoptiles and radicles and a greater radicle number at the germination stage, as well as greater root length, fresh weight, and tiller number per plant at the seedling stage. The yield-related traits of transgenic lines were also improved compared with the wild type, indicating enhanced salt tolerance in transgenic lines overexpressing GmDREB1. Proteomics analysis revealed that osmotic- and oxidative-stress-related proteins were up-regulated in transgenic wheat leaves under salt stress conditions. Transgenic wheat had higher levels of proline and betaine and lower levels of malondialdehyde and relative electrolyte leakage than the wild type. These results suggest that GmDREB1 regulates the expression of osmotic- and oxidative-stress-related proteins that reduce the occurrence of cell injury caused by high salinity, thus improving the salt tolerance of transgenic wheat.
Gray leaf spot (GLS), caused by Cercospora zeae-maydis, is an important foliar disease of maize (Zea mays L.) worldwide, resistance to which is controlled by multiple quantitative trait loci (QTL). To gain insights into the genetic architecture underlying the resistance to this disease, an association mapping population consisting of 161 inbred lines was evaluated for resistance to GLS in a plant pathology nursery at Shenyang in 2010 and 2011. Subsequently, a genome-wide association study, using 41,101 single-nucleotide polymorphisms (SNPs), identified 51 SNPs significantly (P < 0.001) associated with GLS resistance, which could be converted into 31 QTL. In addition, three candidate genes related to plant defense were identified, including nucleotide-binding-site/leucine-rich repeat, receptor-like kinase genes similar to those involved in basal defense. Two genic SNPs, PZE-103142893 and PZE-109119001, associated with GLS resistance in chromosome bins 3.07 and 9.07, can be used for marker-assisted selection (MAS) of GLS resistance. These results provide an important resource for developing molecular markers closely linked with the target trait, enhancing breeding efficiency.
The objective of this study was to investigate whether and how exogenous abscisic acid (ABA) is involved in mediating starch accumulation in the grain and redistribution of carbohydrates during grain filling of two wheat cultivars with different staygreen characteristics. At blooming stage, plants of Wennong 6 (a staygreen cultivar) and Jimai 20 (control) were sprayed with 10 mg L− 1 abscisic acid (ABA) for 3 days. The application of ABA significantly (P < 0.05) increased grain filling rate, starch accumulation rate and content, remobilization of dry matters to kernels, and 1000-grain weight of the two cultivars. Exogenous ABA markedly (P < 0.05) increased grain yield at maturity, and Wennong 6 and Jiami 20 showed 14.14% and 4.86% higher compared yield than the control. Dry matter accumulation after anthesis of Wennong 6 was also significantly (P < 0.05) influenced by exogenous ABA, whereas that of Jimai 20 was unchanged. Application of ABA increased endogenous zeatin riboside (ZR) content 7 days after anthesis (DAA), and spraying ABA significantly increased endogenous indole-3-acetic acid (IAA) and ABA contents from 7 to 21 DAA and decreased gibberellin (GA3) content at 14 DAA, but increased GA3 content from 21 to 35 DAA. The results suggested that increased yield of staygreen was due to greater starch assimilation owing to a higher filling rate and longer grain-filling duration.
Several statistical methods have been developed for analyzing genotype × environment (GE) interactions in crop breeding programs to identify genotypes with high yield and stability performances. Four statistical methods, including joint regression analysis (JRA), additive mean effects and multiplicative interaction (AMMI) analysis, genotype plus GE interaction (GGE) biplot analysis, and yield-stability (YSi) statistic were used to evaluate GE interaction in 20 winter wheat genotypes grown in 24 environments in Iran. The main objective was to evaluate the rank correlations among the four statistical methods in genotype rankings for yield, stability and yield-stability. Three kinds of genotypic ranks (yield ranks, stability ranks, and yield-stability ranks) were determined with each method. The results indicated the presence of GE interaction, suggesting the need for stability analysis. With respect to yield, the genotype rankings by the GGE biplot and AMMI analysis were significantly correlated (P < 0.01). For stability ranking, the rank correlations ranged from 0.53 (GGE-YSi; P < 0.05) to 0.97 (JRA-YSi; P < 0.01). AMMI distance (AMMID) was highly correlated (P < 0.01) with variance of regression deviation (S2di) in JRA (r = 0.83) and Shukla stability variance (σ2) in YSi (r = 0.86), indicating that these stability indices can be used interchangeably. No correlation was found between yield ranks and stability ranks (AMMID, S2di, σ2,22, and GGE stability index), indicating that they measure static stability and accordingly could be used if selection is based primarily on stability. For yield-stability, rank correlation coefficients among the statistical methods varied from 0.64 (JRA-YSi; P < 0.01) to 0.89 (AMMI-YSi; P < 0.01), indicating that AMMI and YSi were closely associated in the genotype ranking for integrating yield with stability performance. Based on the results, it can be concluded that YSi was closely correlated with (i) JRA in ranking genotypes for stability and (ii) AMMI for integrating yield and stability.
The widely used herbicide glyphosate targets 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS). Glyphosate acetyltransferase (GAT) effectively detoxifies glyphosate by N-acetylation. With the aim of identifying a new strategy for development of glyphosate-tolerant crops, the plant expression vector pG2-GAT harboring gat and G2-aroA (encoding EPSPS) has been transformed into tobacco (Nicotiana tabacum) to develop novel plants with higher tolerance to glyphosate. Results from Southern and Western blotting analyses indicated that the target genes were integrated into tobacco chromosomes and expressed effectively at the protein level. Glyphosate tolerance was compared among transgenic tobacco plants containing gat, G2-aroA, or both genes. Plants containing both gat and G2-aroA genes were the most glyphosate-tolerant. This study has shown that a combination of different strategies may result in higher tolerance in transgenic crops, providing a new approach for development of glyphosate-tolerant crops.
Simple sequence repeat (SSR) markers have previously been applied to linkage mapping of the pea (Pisum sativum L.) genome. However, the transferability of existing loci to the molecularly distinct Chinese winter pea gene pool was limited. A novel set of pea SSR markers was accordingly developed. Together with existing SSR sequences, the genome of the G0003973 (winter hardy) × G0005527 (cold sensitive) cross was mapped using 190 F2 individuals. In total, 157 SSR markers were placed in 11 linkage groups with an average interval of 9.7 cM and total coverage of 1518 cM. The novel markers and genetic linkage map will be useful for marker-assisted pea breeding.
