15 May 2014, Volume 26 Issue 3
    

  • Select all
    |
  • WANG Li, MU Chun, ZHANG Ming-Cai, DU Ming-Wei, TIAN Xiao-Li, LI Zhao-Hu
    Cotton Science. 2014, 26(3): 189-196. https://doi.org/10.11963/cs140301
    Abstract ( ) Download PDF ( )   Knowledge map   Save
    Virus-induced gene silencing(VIGS) is a new reverse genetics method used in rapid identification of gene function in plant and has become one of the most widely used plant functional genomics tools. Ent-copalyl diphosphate synthase(CPS) is the first key enzyme involved in gibberellin (GA) biosynthesis. We aimed to elucidate the effect of silencing GhCPS by VIGS on the growth and endogenous hormone content of cotton seedlings. In this study, a 493-bp fragment of GhCPS was amplified from the cotton cultivar(Xinkang 4) cDNA using RT-PCR and was subsequently inserted into the pTRV-RNA2 vector using EcoRⅠand KpnI sites, respectively. pTRV-GhCPS construct was transformed into Agrobacterium tumefaciens GV3101 and was used to transform cotton seedlings at the cotyledon stage with cotton cultivar Xinkang 4 grown in a growth chamber under pot culture. The results showed that the expression level of GhCPS in GhCPS VIGS plants was only 52% of that in pTRV control plants. Silencing GhCPS resulted in 33% and 60% reductions in GA4 content in the 1st and 2nd leaves, respectively, and a slight decrease of IAA content but no obvious changes in ZR and iPA contents in both leaves. ABA content in the 2nd leaf was decreased by 48%. The height of GhCPS VIGS plants was reduced by 64%. The area of the 1st and 2nd leaves in GhCPS VIGS plants was decreased by 26% and 47%, respectively. Silencing GhCPS increased total chlorophyll contents by 29% and 63%, and carotenoids contents by 27% and 46% in the 1st and 2nd leaves, respectively. Net photosynthesis rate, stomatal conductance, intercellular CO2 concentration and transpiration rate in the 1st leaf of GhCPS VIGS plants were enhanced by 23%、40%、9% and 31%, respectively. The results suggest that silencing GhCPS can disturb the balance of endogenous hormone and inhibit the growth of cotton seedlings. The rate of photosynthesis per leaf area can be enhanced by silencing GhCPS.
  • GU Chao, GUO Dan-Li, ZHANG Feng, LI Xue-Yuan, AI Xian-Tao, HUANG Xian-Zhong
    Cotton Science. 2014, 26(3): 197-203. https://doi.org/10.11963/cs140302
    Abstract ( ) Download PDF ( )   Knowledge map   Save
    The phosphatidylethanolamine-binding protein(PEBP) gene family is present in angiosperms with three subfamilies, FT-like, TFL1-like and MFT-like, which having been shown to act as key regulators to control growth and differentiation. Compared with the other two kinds, the exact molecular function of MFT-like subfamily genes is still not clear. One MFT(MOTHER OF FT AND TFL1)-like homolog was isolated from Gossypium barbadense L. cv. Xinhai 14 through RT-PCR and RACE method, which was designated GbMFT2 gene(GenBank accession number: KF739071). The open reading frame(ORF) of GbMFT2 is 519 bp in length, encoding a putative protein of 172 amino acids,containing a PEBP conserved motif. Sequence alignment showed that the nucleotide of GbMFT2 gene has only 56.7% similarity with GbMFT1 gene and the amino acids of GbMFT2 protein has 62.7% similarity with GbMFT1 protein. Phylogenetic analysis revealed that GbMFT2 belong to the MFT-like clade. Expression analysis by qRT-PCR indicated that GbMFT2 displayed a much broader expression range, with a maximum expression in stem and ovule. Expression analysis by semi-quantitative RT-PCR method indicated that GbMFT2 expression levels in germinating seeds were low, but gradually elevated when treated along with increased concentrations of ABA. Our research showed that the expression of GbMFT2 was up-regulated by ABA in germinating seeds, showing significant differences compared with GbMFT1 in the case of ABA treatment.
  • ZHENG Yun-Na, DAI Hua-Qin, CAO Yue-Fen, LI Jin-Juan, RONG Jun-Kang, DING Ming-Quan
    Cotton Science. 2014, 26(3): 204-212. https://doi.org/10.11963/cs140303
    Abstract ( ) Download PDF ( )   Knowledge map   Save
    In order to study the gene regulatory network underlining the cotton fiber initiation, a yeast one hybrid cDNA library of the Gossypium barbadence L. ovule at the stage of flowering(0 DPA, 0 days post anthesis) was constructed and used to screen the potential upstream genes of GbPDF2, an important gene related to fiber initiation, using the Matchmaker Gold Yeast One-Hybrid Library Screening System developed by the Clonetech company. The promoter region of GbPDF2 was cloned and inserted into the vector pAbAi. The recombination bait vector and the cDNA library within the pGADT7-Rec fusion expression vectors were co-transformed into the yeast Y1HGold. The estimated cDNA library storage capacity is almost 2.23 × 106, recombinant rate is 95.8 % and inserted PCR fragments sizes are 300 bp~2000 bp, of which the fragment length of 109 positive insertion clones are larger than 1000 bp. The positive yeast colonies were harvested and cultured for extracting the plasmid, followed by PCR and sequencing analysis. Several transcription factors are screened out such as WRKY7 transcription factor, WD-repeat protein 40, IAA responsible transcription factors. This research has laid the fundament in applying yeast one hybrid method to study signal transduction pathway of fiber initiation in cotton.
  • MA Ling-Ling, WEI Yan-Hong, HE Lan-Lan, CHAI Meng-Liang, ZHU Hua-Guo, SUN Jie, ZHANG Wei
    Cotton Science. 2014, 26(3): 213-220. https://doi.org/10.11963/cs140304
    Abstract ( ) Download PDF ( )   Knowledge map   Save
    In this study, biolistics transformation system was established with apical meristems of Xinhai No.13, a variety of Gossypium barbadense L. cultured in Xinjiang. Several factors of the system are studied, which contained the wrapped concentration of DNA, the type of microcarrier, the early penetration processing time, bombardment condition and screening technique, et al. The optimal transformation system is as follows under other constant conditions. Microcarriers of a diameter of 1.0 μm were covered up with the DNA concentration of 1.5 μg·μL-1. Apical meristems were bombarded with the helium pressure of 7.584MPa(1100 psi), and with the bombardment distance of 6 cm. Target materials were cultured for 12 h before being bombarded and 16 h after being bombarded in the osmotic medium. After recovering for 24 h in the recovery medium, target materials were moved to the kanamycin screening medium containing the concentration of 120 mg·L-1 kanamycin to screen directly for 35 days. Resistant seedlings were induced to take roots in the induction rooting medium adding 4 g·L-1 activated charcoal granule and the regenerating plants were obtained. After PCR and RT-PCR detection, five transgenic regenerated plants were obtained in the end in which the gene EPSPS were expressed.
  • WANG Xiao-Nan, QI Hong, ZHANG Jin-Feng, GUANG Yang-Qi, TANG Can-Ming
    Cotton Science. 2014, 26(3): 221-227. https://doi.org/10.11963/cs140305
    Abstract ( ) Download PDF ( )   Knowledge map   Save
    In order to analyse the infection mechanism of V. dahliae, sGFP encoding green fluorescent protein was transferred into defoliating V.dahliae isolate VD07038 and mCherryRFP encoding red fluorescent protein was transferred into non-defoliating V.dahliae Bp2 via Agrobacterium tumefaciens-mediated transformation. Finally, positive transformants tagged by green fluorescent protein or red fluorescent protein were obtained. The hygromycin B genetic stability of transgenic progenies was tested by series culture and molecular verification. According to colony morphology, growth rate and pathogenicity, we found that traits of most of transformants were similar to those of wild-type, few transformants tended to form mutants. Tansformants Bp2R-30 did not produce microsclerotia and its pathogenicity decreased significantly. Using the GFP-tagged V. dahliae VD07038G-10 isolate, colonization of the fungus in roots of the susceptible cotton Sumian 22 was investigated. As the results showed that, 12 hours after inoculation via the root dipping methods, roots surface of Sumian 22 were covered with conidia, 7-9 days after inoculation, hyphae from the leading edge of the colony progressed acropetally up the xylem vessels of infected roots. V. dahliae VD07038G-10 tagged by fluorescent protein can be used for observation its infection process in real time on cotton roots and identification of resistance of different cotton varieties to this strain. This study provide a new method for resistance evaluation of V. dahliae.
  • LI Na, ZHANG Shuai, LUO Jun-Yu, 吕Li-Min , CUI Jin-Jie, LI Guo-Qing
    Cotton Science. 2014, 26(3): 228-236. https://doi.org/10.11963/cs140306
    Abstract ( ) Download PDF ( )   Knowledge map   Save
    Chemosensory proteins(CSPs), which belongs to a class of small soluble polypeptides in insect chemical receptors, may be related with the process of storing and releasing the chemical odors for insects. In this study, one full-length cDNA of chemosensory protein gene was cloned from Aphis gossypii, named AgosCSP2 (GenBank accession No. KC017751). The temporal expression pattern of the gene was studied by qRT-PCR analysis. The results indicated that the expression levels in nymph were significantly higher than those in adult; the AgosCSP2 was higher expressed in the host of cotton than that in the overwintering host hibiscus and it was higher expressed in leg than that in other organizations. The binding characteristics showed that AgosCSP2 had low binding abilities with odorant ligands. The result suggested that the AgosCSP2 gene most likely functioned in the process of cotton aphid nymph adaptation to host, but not functioned as the chemosensory of the insect. This experiment may be valuable for studying the role of AgosCSP2 in A. gossypii chemical sensors.
  • ZHANG Zheng-Zhuan, TANG Lei-Peng, WANG Yu, HE Jun-Guang, CAI Wen-Long, RONG Jun-Kang, LI Fei-Fei
    Cotton Science. 2014, 26(3): 237-243. https://doi.org/10.11963/cs140307
    Abstract ( ) Download PDF ( )   Knowledge map   Save
    For producing the glyphosate-resistant cotton plants, we developed an Agrobacterium-mediated transformation system from embryogenic calli(EC) in which glyphosate was used as a selective agent. In this study, glyphosate-resistant gene aroA was transformed to embryogenic calli(EC) of Gossypium hirsutum L. cv. WC and YZ-1. And the calli were transferred onto culture medium, respectively, with 5 and 10 mg·L-1 glyphosate for selection. After two or three cycles of glyphosate-resistant screening, resistant EC was obtained and transgenic plantlets were regenerated from embryoid and grafted in greenhouse. Glyphosate-resistant positive plants were identified by PCR analysis and glyphosate-resistant screening. In this study, 33 and 10 glyphosate-resistant positive plants were obtained from WC and YZ-1, respectively, and the corresponding transformation efficiency was 82.5% and 62.5%. This transformation system provides not only an efficient method for cotton genetic transformation but also important germplasm resources for cotton breeding. Meanwhile, the selection marker gene aroA can be transformed into cotton genome together with other target gene for functional analysis.
  • HUANG Chun-Yan, WANG Deng-Wei, XIAO Li-Juan, WANG Ya-Fang
    Cotton Science. 2014, 26(3): 244-251. https://doi.org/10.11963/cs140308
    Abstract ( ) Download PDF ( )   Knowledge map   Save
    Hyperspectral reflectance data were recorded by the ASD portable non-imaging hyperspectral spectrometer at five growth stages of cotton in an experimental field including two cotton cultivars, Xinluzao No.33, No.13, with four water treatments and cotton canopy thermal images were taken with a Fluke radiometric infrared video camera, thermal images can be used to extract canopy sunlit leaves temperature(TL). Meanwhile, hyperspectral data were processed to calculate the normalized difference vegetation index(NDVI), ratio vegetative index(RVI), reflectance at red spectrum 620 nm and near-infrared 850 nm wavelength(ρ620, ρ850), respectively. The results showed that canopy leaves temperature(TL) of two cotton cultivars with four water treatments reached the higher value at full flowering stage, full flowering and boll-forming stage, their maximum value were both at full boll stage, the lower TL both occurred at flowering stage and initial open boll stage. Cotton reflectance were reduced at near-infrared and increased at red spectrum wavelength, corresponding NDVI and RVI were less, TL was greater in water stressed plots. The spectrum reflectance at red and NIR wavelength, NDVI, RVI and TL were opposite variation at full water plots than these parameters at water stress treatments. The relationships between NDVI, RVI and TL(rNDVI-TL= -0.4518**,rRVI-TL= -0.5274**,n=38, α=1%)were both stronger than the relationships between ρ620, ρ850 and TL, respectively. It showed that combination of infrared thermography and hyperspectral remote sensing technology have potential to rapidly and nondestructively monitor water status of cotton.
  • GUO Bao-Sheng, WANG Kai-Hui, LIU Su-恩, ZHAO Cun-Peng, GENG Jun-Yi, ZHANG Xiang-Yun, HUA Jin-Ping
    Cotton Science. 2014, 26(3): 252-259. https://doi.org/10.11963/cs140309
    Abstract ( ) Download PDF ( )   Knowledge map   Save
    This article outlined the progresses in the research on creation of resistance germplasm to Verticillium wilt in Gossypium hirsutum, and the development and achievement in resistant varieties breeding, as well as primitive understanding on the inheritance of cotton resistance to the disease and discovery of resistance genes. Distant hybridization breeding served as a powerful way to create the germplasm resistance to Verticillium wilt in upland cotton. Genetic engineering, a new technology, can be employed in the project of germplasm creation. Because of the complexity of the resistant mechanism, it seems that the resistance was not changed by transferring one or two major effect genes in resistant germplasm breeding. Resistance genes and pathogenesis-related genes discovered from upland cotton or relative species, the revealing of the mechanism and its defense will provide theoretical support for molecular design breeding strategy.
  • LIN Ling, ZHANG Xin, DENG Sheng
    Cotton Science. 2014, 26(3): 260-267. https://doi.org/10.11963/cs140310
    Abstract ( ) Download PDF ( )   Knowledge map   Save
    Cotton Verticillium wilt caused by Verticillium dahliae Kleb. has become one of the main impediments to cotton sustainable production in China in recent years.This disease is a soil-borne and seed-borne vascular disease and difficult to control. The host range of V. dahliae is unusually wide as a plant pathogen.This paper summarized the latest research progresses of the pathogenicity differentiation, genome sequence, pathogenesis mechanism, microsclerotial development and germination mechanism of V. dahliae, molecular mechanism of disease resistance in cotton as well as the control methods of cotton Verticillium wilt.
  • ZHANG Wen-Wei, LI Sha, JIAN Gui-Liang, SI Ning, QI Fang-Jun
    Cotton Science. 2014, 26(3): 268-273. https://doi.org/10.11963/cs140311
    Abstract ( ) Download PDF ( )   Knowledge map   Save
    17 Alternaria isolates were obtained from two municipalities,an autonomous region and four provinces in China. To clarify the species of them, both observation on morphological characteristics and ITS sequencing were used. The results showed that 16 isolates including A1 etc. were identified as Alternaria alternata, and A3 isolate was identified as Alternaria macrospore.
  • ZHANG Yan, DING Ze-Guo, WANG Sheng-Fen, ZHANG Gui-Rong, LI Zhi-Kun, ZHANG Gui-Yin, WU Li-Qiang, MA Zhi-Ying
    Cotton Science. 2014, 26(3): 274-278. https://doi.org/10.11963/cs140312
    Abstract ( ) Download PDF ( )   Knowledge map   Save
     A full-length cDNA library inoculated by Verticillium dahliae was constructed with the Gossypium barbadense cv. Pima 90-53 roots based on SMART technique, the quality evaluatation and EST analysis of this library were finished. The recombinant rate of present library was 92.3%,the titer was 1.1×106 pfu·mL-1,and the average size of inserted cDNA fragments was about 1.5 kb. Bioinformatics analysis of EST data indicated that a lot of unigenes with homology to known defense-related genes in other plants were found in our library. Clusters of orthologous group(COG) functional classification found that more genes which involved in signal transduction, defense mechanisms, cytoskeleton and secondary metabolites biosynthesis/transport/catabolism were detected. And the highly-expressed genes were detected, which mainly contained pathogenesis-related proteins(PR proteins), glutathione-S-transferase(GST), cyclophilin, short chain alcohol dehydrogenase, ascorbic acid peroxidase, transcription factor, and so on. These will be of significance to understanding the molecular mechanism of cotton Verticillium wilt resistance.
  • LI Jie, ZONG Tao, LIU Xiang-Ying, BAI Lian-Yang
    Cotton Science. 2014, 26(3): 279-282. https://doi.org/10.11963/cs140313
    Abstract ( ) Download PDF ( )   Knowledge map   Save
    In order to evaluate the resistance of Eleusine indica populations to haloxyfop-R-methyl, whole-plant bioassay was measured, and the differences on GST of the resistant and susceptible populations to haloxyfop-R-methyl were preliminarily determined. The results of whole-plant bioassay showed that populations of Eleusine indica from the collecting sites of Hunan Province developed different levels of resistance to haloxyfop, and the highest resistant index(RI) was 18.4, which was found in populations from Hanshou. The next were that of Datonghu and Xiangyin population. The GST activity of resistant population decreased immediately and then increased rapidly at 5 days after treatment, with the peak value 9 days after, while that of susceptible population declined at first and then increased, then came down to the control value 9 days later. There were great difference between GST activities of Hanshou (resistant) and Dingcheng (susceptible) population, which indicated difference of GST metabolizing capacity is an important factor for Eleusine indica's resistance to haloxyfop-R-methyl.