Small RNAs (sRNAs), the key components of RNA interference (RNAi) or RNA silencing, can mediate cell-autonomous gene silencing and function as signaling molecules across species. Microbe-induced gene silencing (MIGS), which is based on interspecies RNAi, is an effective approach for controlling fungal diseases in crops. The enolase gene VdEno is essential for the growth and development of the fungal pathogen Verticillium dahliae, which causes cotton Verticillium wilt. In this study, we engineered Trichoderma harzianum (Th) to express the double-stranded RNA (dsRNA) targeting VdEno. The engineered strain Th-VdEnoi successfully generated VdEno-specific small interfering RNA (siVdEno). We further confirmed that Th-VdEnoi effectively induced VdEno silencing at the translational level. The results of crop protection assays revealed that the cotton plants co-inoculated with V. dahliae (strain V592) and Th-VdEnoi presented significantly reduced disease severity and lower fungal biomass in their roots than the control plants inoculated with V. dahliae alone or with V. dahliae and Th-GFPi (a control strain expressing GFP-targeting dsRNA). Collectively, our findings demonstrate that VdEno is an effective target for controlling cotton Verticillium wilt and confirm that MIGS is a promising strategy for managing soil-borne fungal pathogens in crops. MIGS provides strong technical support for reducing the application of conventional chemical pesticides, developing eco-friendly biopesticides, and facilitating the sustainable development of agriculture.
Gossypium/microbiology* ; Plant Diseases/prevention & control* ; Gene Silencing ; Ascomycota/genetics* ; RNA Interference ; RNA, Double-Stranded/genetics* ; Hypocreales/genetics* ; RNA, Small Interfering/genetics* ; Verticillium/genetics* ; Fungal Proteins/genetics*

Plant trichomes are special structures that originate from epidermal outgrowths. Trichomes play an important role in plant defense against pests and diseases, and possess economic and medicinal values. Study on molecular mechanism of plant trichomes will contribute to the molecular design breeding and genetic improvement of crops. In recent years, the regulation mechanism of trichome development has been basically clarified in the model plant Arabidopsis thaliana, while great progresses are also found in other plant species. In this review, we focus on the developmental regulation of trichome formation from gene and phytohormones levels in Arabidopsis and cotton (with unicellular trichomes), as well as in tomato and Artemisia annua (with multicellular trichomes). The research progress associated with trichomes is also introduced in other typical monocotyledons and dicotyledons. Finally, the research and application of plant trichomes are prospected.
Arabidopsis/genetics* ; Gene Expression Regulation, Plant ; Gossypium/genetics* ; Lycopersicon esculentum ; Plant Growth Regulators/metabolism* ; Trichomes/genetics*

Arabidopsis ; genetics ; growth & development ; microbiology ; Gossypium ; growth & development ; microbiology ; Green Fluorescent Proteins ; Hyphae ; growth & development ; pathogenicity ; Plant Diseases ; genetics ; microbiology ; Plant Roots ; microbiology ; Spores, Fungal ; growth & development ; pathogenicity ; Verticillium ; growth & development ; pathogenicity

Cloning of flanking sequences of double-copy gene is a challenge in molecular biology. We developed a method to solve this problem by combining an optimized inverse PCR (iPCR) with TAIL-PCR. First, Southern blotting analysis was used to determine a proper restriction enzyme that could obtain proper-length restriction fragments that contained the target gene. Then optimized iPCR was performed to amplify the restriction fragments that contained the separated copies of the gene. Based on the obtained sequences, TAIL-PCR was performed to amplify further flanking regions of the gene. With this method, we obtained all of the EcoR I restriction fragments (2.2-5.1 kb) and Hind III restriction fragments (8.5-11.7 kb) of mitochondrial atpA gene in cytoplasmic male sterile (CMS) line and maintainer line of Upland cotton. The results showed that this method was an efficient approach to clone flanking sequences of double-copy gene.
Chromosome Walking ; Cloning, Molecular ; Gene Expression Regulation, Plant ; Genes, Mitochondrial ; Genes, Plant ; genetics ; Gossypium ; genetics ; Plant Proteins ; genetics ; metabolism ; Polymerase Chain Reaction ; methods ; Terminal Repeat Sequences

Aquaporin belongs to a highly conserved group of membrane proteins called major intrinsic proteins (MIPs) that facilitate water transport across biological membranes. Aquaporins are membrane water channels that play critical roles in controlling the water content of cells and tissues. We focused on GhPIP1;2 which belongs to the PIP subfamily and GhgammaTIP1 which belongs to the gammaTIP group of the TIP subfamily. Northern blot analysis with gene-specific probes and real-time PCR demonstrated that GhPIP1;2 and GhgammaTIP1 are predominantly expressed during cotton fiber elongation, with the highest expression levels at 5 days post anthesis. The high and preferential expression of GhPIP1;2 and GhgammaTIP1 suggests that they may play important roles in supporting the rapid influx of water into vacuoles during cotton fiber cell expansion. Also, the effects of Ca2+ on aquaporins in salinity-stressed plants were studied. Researchers treated the protoplasts and plasma membrane with NaCl or CaCl2, alone or in combination. Under saline conditions, osmotic water permeability (Pf) values decreased in protoplasts and plasma membrane vesicles, and the same reduction was observed in the PIP1 aquaporin abundance, indicating inhibitory effects of NaCl on aquaporin functionality and protein abundance. Two different actions of Ca2+ were observed. Increase in free cytosolic calcium concentrations associated with stress perception may lead to aquaporin closure, however, the extra-calcium would lead to an upregulation of aquaporins. Meanwhile, experiments have demonstrated HvPIP2;1, one of barley aquaporins, has a higher water and CO2 transport activity. The goal of our plant aquaporin research is to determine the key aquaporin species responsible for water and CO2 transport, and to improve plant water relations, stress tolerance, CO2 uptake or assimilation, and plant productivity.
Aquaporins ; physiology ; Cell Enlargement ; Cotton Fiber ; Gossypium ; metabolism ; physiology ; Photosynthesis ; physiology ; Plant Proteins ; physiology ; Sodium Chloride ; pharmacology ; Stress, Physiological ; physiology

One hundred and sixty-eight genotypes of cotton from the same growing region were used as a germplasm group to study the validity of different genetic distances in constructing cotton core subset. Mixed linear model approach was employed to unbiasedly predict genotypic values of 20 traits for eliminating the environmental effect. Six commonly used genetic distances (Euclidean, standardized Euclidean, Mahalanobis, city block, cosine and correlation distances) combining four commonly used hierarchical cluster methods (single distance, complete distance, unweighted pair-group average and Ward's methods) were used in the least distance stepwise sampling (LDSS) method for constructing different core subsets. The analyses of variance (ANOVA) of different evaluating parameters showed that the validities of cosine and correlation distances were inferior to those of Euclidean, standardized Euclidean, Mahalanobis and city block distances. Standardized Euclidean distance was slightly more effective than Euclidean, Mahalanobis and city block distances. The principal analysis validated standardized Euclidean distance in the course of constructing practical core subsets. The covariance matrix of accessions might be ill-conditioned when Mahalanobis distance was used to calculate genetic distance at low sampling percentages, which led to bias in small-sized core subset construction. The standardized Euclidean distance is recommended in core subset construction with LDSS method.
Cluster Analysis ; Genetic Variation ; Genotype ; Gossypium ; genetics ; Linear Models ; Principal Component Analysis

The open reading frame of Spinacia oleracea Betaine Aldehyde Dehydrogenase (SoBADH) was retrieved from Spinacia oleracea and inserted into the Agrobacterium tumefaciens binary vector pBin438, which was driven by CaMV35S promoter, and produced the new binary vector pBSB. A. tumefaciens LBA4404 carrying this plasmid was used in genetic transformation of plants. Forty-five primary transgenic plants were detected by PCR and verified by the Southern blotting from 65 regenerated plants, of which 27 transgenic plants had only one copy of T-DNA. The Northern blotting and Western blotting analysis indicated that the SoBADH gene had been transcribed mRNA and expression protein in the transgenic cotton lines. The testing of SoBADH activity of transgenic plant leaves showed that the enzyme activity was much higher than that of the non-transgenic cotton. The growth of transgenic plants was well under the salinity and freezing stress, whereas the non-transgenic plant grew poorly and even died. Challenging with salinity, the height and fresh weight of transgenic plants was higher compared with those of non-transgenic plants. Under the freezing stress, the relative conductivity of leaf electrolyte leakage of the transgenic cotton lines was lower than that of non-transgenic plants. These results demonstrated that the SoBADH gene could over express in the exogenous plants, and could be used in genetic engineering for cotton stress resistance.
Adaptation, Physiological ; Betaine-Aldehyde Dehydrogenase ; biosynthesis ; genetics ; Cold Temperature ; Gossypium ; enzymology ; genetics ; Plants, Genetically Modified ; enzymology ; genetics ; Salinity ; Spinacia oleracea ; enzymology ; genetics ; Stress, Physiological ; genetics

Virus isolate G6 was obtained from Hibiscus rosa-sinensis showing yellow and leaf curl symptoms in Guangzhou, Guangdong Province. The complete nucleotide sequence of DNA-A was determined to be 2 737 nucleotides encoding six potential ORFs. Comparison showed that G6 DNA-A had more than 89% sequence identify with all isolates of Cotton leaf curl Multan virus (CLCuMV) and shared the highest sequence identify (96.1%) with CLCuMV isolate 62. G6 DNA-A had 87.1%-89.8% sequence identity with those of CLCuRV isolates, while less than 87% identities with other begomoviruses. Phylogenetic analysis of G6 DNA-A and selected begomoviruses showed that G6 was most closely related to CLCuMV isolates, and they clustered together as a separate branch. Satellite DNA molecule (G6 DNAbeta) was found to be associated with G6 using the primers beta01 and beta02. G6 DNAbeta contains 1346 nucleotides, with a potential functional ORF (C1) in complementary sense DNA. Pairwise comparison indicated that G6 DNAbeta had the highest sequence identities with CLCuMV DNAbeta (92.1%) and CLCuRV DNAbeta (88.7%), but less than 80% sequence identities with other reported satellite DNA molecules. Phylogenetic analysis indicated that G6 DNAbeta was most closely related to CLCuMV DNAbeta and the two DNAbetas clustered together as a separate branch, and formed the main branch with DNAbeta of CLCuRV and MYVV-Y47. It is concluded that G6 infecting Hibiscus rosa-sinensis is an isolate of CLCuMV.
Base Sequence ; DNA, Satellite ; chemistry ; DNA, Viral ; chemistry ; Geminiviridae ; classification ; genetics ; Gossypium ; virology ; Hibiscus ; virology ; Phylogeny

OBJECTIVETo compare the difference of the therapeutic effects between Yang's cotton moxibustion and hormonotherapy on neurodermatitis.

METHODSSixty cases of neurodermatitis were divided into a treatment group and a control group according to random number table. The treatment group were treated with Yang's cotton moxibustion and the control group with triamcinolone and urea cream. After treatment for one week, the therapeutic effect was start to be observed. After the treatment was given for 4 weeks, the therapeutic effect was evaluated.

RESULTSThe total effective rate was 93.3% in the treatment group and 80.0% in the control group, with a significant difference between the two groups (P < 0.05).

CONCLUSIONYang's cotton moxibustion has an obvious therapeutic effect on neurodermatitis with a good safety.

Adolescent ; Adult ; Aged ; Female ; Gossypium ; Humans ; Male ; Middle Aged ; Moxibustion ; methods ; Neurodermatitis ; therapy

Wild cotton species can contribute a valuable gene pool for agronomically desirable cultivated tetraploid cultivars. In order to exploit diploid cotton a regeneration system is required to achieve transformation based goals. The present studies aimed at optimizing the conditions for regeneration of local varieties as well as wild species of cotton. Different callus induction media were tested with varying concentrations of hormones in which sucrose was used as nutritional source. Different explants (hypocotyls, cotyledon, root) were used to check the regeneration of both local cotton plants and wild relatives using T & G medium, BAP medium, CIM medium, EMMS medium, and cell suspension medium. Different stages of embryogenicity such as early torpedo stage, late torpedo stage, heart stage, globular stage and cotyledonary stage were observed in wild relatives of cotton. The results of this study pave the way for establishing future transformation methods.
2,4-Dichlorophenoxyacetic Acid ; Benzyl Compounds ; Cotyledon ; growth & development ; Culture Media ; Gossypium ; embryology ; genetics ; growth & development ; metabolism ; Hypocotyl ; growth & development ; Kinetin ; Naphthaleneacetic Acids ; Plant Growth Regulators ; Purines ; Regeneration ; physiology ; Transformation, Genetic ; Zeatin