Genome-Wide Identification and Functional Analysis of RF2 Gene Family and the Critical Role of GhRF2-32 in Response to Drought Stress in Cotton.

Cotton is an important natural fiber crop. The RF2 gene family is a member of the bZIP transcription factor superfamily, which plays an important role in plant resistance to environmental stresses. In this paper, the RF2 gene family of four cotton species was analyzed genome-wide, and the key gene RF2-32 was cloned for functional verification. A total of 113 RF2 genes were identified in the four cotton species, and the RF2 family was relatively conserved during the evolution of cotton. Chromosome mapping and collinear analysis indicated that fragment replication was the main expansion mode of RF2 gene family during evolution. Cis-element analysis showed that there were many elements related to light response, hormone response and abiotic stress response in the promoters of RF2 genes. The transcriptome and qRT-PCR analysis of RF2 family genes in upland cotton showed that RF2 family genes responded to salt stress and drought stress. GhRF2-32 protein was localized in the cell nucleus. Silencing the GhRF2-32 gene showed less leaf wilting and increased total antioxidant capacity under drought and salt stress, decreased malondialdehyde content and increased drought and salt tolerance. This study revealed the evolutionary and functional diversity of the RF2 gene family, which laid a foundation for the further study of stress-resistant genes in cotton.

Effects of Single and Combined Drought and Salinity Stress on the Root Morphological Characteristics and Root Hydraulic Conductivity of Different Winter Wheat Varieties.

Water shortages and crop responses to drought and salt stress are related to the efficient use of water resources and are closely related to food security. In addition, PEG or NaCl stress alone affect the root hydraulic conductivity (Lpr). However, the effects of combined PEG and NaCl stress on Lpr and the differences among wheat varieties are unknown. We investigated the effects of combined PEG and NaCl stress on the root parameters, nitrogen (N) and carbon content, antioxidant enzymes, osmotic adjustment, changes in sodium and potassium, and root hydraulic conductivity of Yannong 1212, Heng 4399, and Xinmai 19. PEG and NaCl stress appreciably decreased the root length (RL), root surface area (RS), root volume (RV), K+ and N content in shoots and roots, and Lpr of the three wheat varieties, while the antioxidant enzyme activity, malondialdehyde (MDA), osmotic adjustment, nonstructural carbon and Na+ content in shoots and roots, etc., remarkably remained increased. Furthermore, the root hydraulic conductivity had the greatest positive association with traits such as RL, RS, and N and K+ content in the shoots of the three wheat varieties. Moreover, the RL/RS directly and actively determined the Lpr, and it had an extremely positive effect on the N content in the shoots of wheat seedlings. Collectively, most of the root characteristics in the wheat seedlings decreased under stress conditions, resulting in a reduction in Lpr. As a result, the ability to transport nutrients-especially N-from the roots to the shoots was affected. Therefore, our study provides a novel insight into the physiological mechanisms of Lpr.

Genome-Wide Identification and Characterization of the PPO Gene Family in Cotton (Gossypium) and Their Expression Variations Responding to Verticillium Wilt Infection.

Polyphenol oxidases (PPOs) are copper-binding metalloproteinases encoded by nuclear genes, ubiquitously existing in the plastids of microorganisms, plants, and animals. As one of the important defense enzymes, PPOs have been reported to participate in the resistant processes that respond to diseases and insect pests in multiple plant species. However, PPO gene identification and characterization in cotton and their expression patterns under Verticillium wilt (VW) treatment have not been clearly studied. In this study, 7, 8, 14, and 16 PPO genes were separately identified from Gossypium arboreum, G. raimondii, G. hirsutum, and G. barbadense, respectively, which were distributed within 23 chromosomes, though mainly gathered in chromosome 6. The phylogenetic tree manifested that all the PPOs from four cotton species and 14 other plants were divided into seven groups, and the analyses of the conserved motifs and nucleotide sequences showed highly similar characteristics of the gene structure and domains in the cotton PPO genes. The dramatically expressed differences were observed among the different organs at various stages of growth and development or under the diverse stresses referred to in the published RNA-seq data. Quantitative real-time PCR (qRT-PCR) experiments were also performed on the GhPPO genes in the roots, stems, and leaves of VW-resistant MBI8255 and VW-susceptible CCRI36 infected with Verticillium dahliae V991, proving the strong correlation between PPO activity and VW resistance. A comprehensive analysis conducted on cotton PPO genes contributes to the screening of the candidate genes for subsequent biological function studies, which is also of great significance for the in-depth understanding of the molecular genetic basis of cotton resistance to VW.

Conservation and Divergence of Phosphoenolpyruvate Carboxylase Gene Family in Cotton.

Phosphoenolpyruvate carboxylase (PEPC) is an important enzyme in plants, which regulates carbon flow through the TCA cycle and controls protein and oil biosynthesis. Although it is important, there is little research on PEPC in cotton, the most important fiber crop in the world. In this study, a total of 125 PEPCs were identified in 15 Gossypium genomes. All PEPC genes in cotton are divided into six groups and each group generally contains one PEPC member in each diploid cotton and two in each tetraploid cotton. This suggests that PEPC genes already existed in cotton before their divergence. There are additional PEPC sub-groups in other plant species, suggesting the different evolution and natural selection during different plant evolution. PEPC genes were independently evolved in each cotton sub-genome. During cotton domestication and evolution, certain PEPC genes were lost and new ones were born to face the new environmental changes and human being needs. The comprehensive analysis of collinearity events and selection pressure shows that genome-wide duplication and fragment duplication are the main methods for the expansion of the PEPC family, and they continue to undergo purification selection during the evolutionary process. PEPC genes were widely expressed with temporal and spatial patterns. The expression patterns of PEPC genes were similar in G. hirsutum and G. barbadense with a slight difference. PEPC2A and 2D were highly expressed in cotton reproductive tissues, including ovule and fiber at all tested developmental stages in both cultivated cottons. However, PEPC1A and 1D were dominantly expressed in vegetative tissues. Abiotic stress also induced the aberrant expression of PEPC genes, in which PEPC1 was induced by both chilling and salinity stresses while PEPC5 was induced by chilling and drought stresses. Each pair (A and D) of PEPC genes showed the similar response to cotton development and different abiotic stress, suggesting the similar function of these PEPCs no matter their origination from A or D sub-genome. However, some divergence was also observed among their origination, such as PEPC5D was induced but PEPC5A was inhibited in G. barbadense during drought treatment, suggesting that a different organized PEPC gene may evolve different functions during cotton evolution. During cotton polyploidization, the homologues genes may refunction and play different roles in different situations.

Identification of the Golden-2-like transcription factors gene family in Gossypium hirsutum.

BACKGROUND: Golden2-Like (GLK) transcription factors are a type of transcriptional regulator in plants. They play a pivotal role in the plant physiological activity process and abiotic stress response. METHODS: In this study, the potential function of GLK family genes in Gossypium hirsutum was studied based on genomic identification, phylogenetic analysis, chromosome mapping and cis-regulatory elements prediction. Gene expression of nine key genes were analyzed by qRT-PCR experiments. RESULTS: Herein, we identified a total of 146 GhGLK genes in Gossypium hirsutum, which were unevenly distributed on each of the chromosomes. There were significant differences in the number and location of genes between the At sub-genome and the Dt sub-genome. According to the phylogenetic analysis, they were divided into ten subgroups, each of which had very similar number and structure of exons and introns. Some cis-regulatory elements were identified through promoter analysis, including five types of elements related to abiotic stress response, five types of elements related to phytohormone and five types of elements involved in growth and development. Based on public transcriptome data analysis, we identified nine key GhGLKs involved in salt, cold, and drought stress. The qRT-PCR results showed that these genes had different expression patterns under these stress conditions, suggesting that GhGLK genes played an important role in abiotic stress response. This study laid a theoretical foundation for the screening and functional verification of genes related to stress resistance of GLK gene family in cotton.

Enhancing the systems productivity and water use efficiency through coordinated soil water sharing and compensation in strip-intercropping.

In arid areas, water shortage is threating agricultural sustainability, and strip-intercropping may serve as a strategy to alleviate the challenge. Here we show that strip-intercropping enhances the spatial distributions of soil water across the 0-110 cm rooting zones, improves the coordination of soil water sharing during the co-growth period, and provides compensatory effect for available soil water. In a three-year (2009-2011) experiment, shorter-season pea (Pisum sativum L.) was sown in alternate strips with longer-season maize (Zea mays L.) without or with an artificially-inserted root barrier (a solid plastic sheet) between the strips. The intercropped pea used soil water mostly in the top 20-cm layers, whereas maize plants were able to absorb water from deeper-layers of the neighboring pea strips. After pea harvest, the intercropped maize obtained compensatory soil water from the pea strips. The pea-maize intercropping without the root barrier increased grain yield by 25% and enhanced water use efficiency by 24% compared with the intercropping with the root barrier. The improvement in crop yield and water use efficiency was partly attributable to the coordinated soil water sharing between the inter-strips and the compensatory effect from the early-maturing pea to the late-maturing maize.