15 July 2012, Volume 24 Issue 4

    

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  Molecular Cloning of Gh14-3-3L2 and Identification of its Protein Interactions in Upland Cotton

  HUI Ying, WANG Jin, SUN Jing, LIU Kang, TANG Can-Ming

  Cotton Science. 2012, 24(4): 285-292.

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  From an EST sequence identified by previous proteomic analysis in our lab we cloned a full-length cDNA, named Gh14-3-3L2, which encodes a cotton(Gossypium hirsutum) 14-3-3 protein that probably plays a regulatory role in cotton fiber initiation and elongation. To unravel the target gene networks of this protein, the interactome of Gh14-3-3L2 was studied using the yeast two-hybrid system and a Pull-down assay. However, this protein was severely toxic to yeast. Thus, this paper focuses on the Pull-down results. Six × His tagged Gh14-3-3L2 was prokaryotically expressed in E. coli and purified for use as bait protein. Gh14-3-3L2 associated prey proteins were isolated and enriched from mixed protein samples derived from ovules 3, 1 or 0 days before anthesis and fibers 3 or 6 days after anthesis of wild-type upland cotton cv. Xuzhou142, the lintless mutant Xuzhou142, and the Ligon lintless 1 mutant using Pull-down technology. The resulting proteins were further isolated by 2-DE and identified by MALDI-MS/MS analysis. Seven Gh14-3-3L2 associated proteins were identified. The putative functions of these proteins included molecular chaperoning, vacuolar trafficking, metabolism, and signal transduction. Some of these proteins, including annexin, vacuolar H+-ATPase and Ran GTPase 3, have been previously linked with fiber development. Our study provides novel data for further elucidation of the molecular functions of Gh14-3-3L2 and the regulatory mechanisms governing cotton fiber development.
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  Study on the Dwarf Mechanism of an Upland Cotton(G. hirsutum L.) Dwarf Mutant, Ari1327

  WANG Xin-Kun, SUN Jun-Ling, PAN Zhao-E, ZHANG Chao, HE Shou-Pu, JIA Yin-Hua, ZHOU Zhong-Li, TANG Can-Ming, DU Xiong-Ming

  Cotton Science. 2012, 24(4): 293-302.

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  Plant height is an important agronomic trait in upland cotton. Ari1327 is a dwarf mutant derived from an American upland cotton variety, Ari971, by ⁶⁰Co γ-ray irradiation. Ari1327 exhibits the dwarf trait during the germination and cotyledon periods. During the budding period, the plant height, internode number and internode length of Ari1327 were significantly(P < 0.01) smaller or fewer than in the wild type. Therefore, the mutant dwarfism was due to the decrease of average internode length. The longitudinal cell length of mutant internodes was significantly shorter than in the wild type(P < 0.01). The difference in transverse cell diameter between the mutant and the wild type was not significant. Thus, the shorter internodes were induced by inhibition of internode cells. The IAA, GA₃ and ABA content of Ari1327 was higher than in the wild type. The height of Ari1327 plants could be restored to wild type levels by exogenous GA₃ application. Exogenous BR and IAA had no effect on Ari1327. Due to over-expression of genes in the IAA synthetic pathway, the IAA content in Ari1327 was remarkably increased. High-density IAA inhibits plant growth; thus, the plant exhibits a dwarf phenotype.
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  Effects of Nitrogen Fertilization on Bt Cotton Growth and Bt Protein Concentration in Leaves under Salinity Stress

  DAI Jian-Long, DONG He-Zhong, DUAN Liu-Sheng, LI Zhen-Huai, LU He-Quan

  Cotton Science. 2012, 24(4): 303-311.

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  It is well known that salinity stress inhibits plant growth and Bt protein synthesis in cotton(Gossypium hirsutum L.), but it is still not clear if fertilization alleviates the inhibitory effects. Two experiments with pot culture and hydroponic culture were conducted using Bt cotton strain K638 in a rain shelter and a greenhouse, respectively, to study the effects of N fertilization and its forms(NO₃^--N and NH₄⁺-N) on plant growth, nitrogen uptake, Bt protein content and the ratio of N in Bt protein to total N under salinity stress. In the pot experiment, a significant interaction was detected between N fertilization and salinity stress on leaf Bt protein content. Under non-saline and low saline(total salt content≈0.15%) soils, N fertilization enhanced plant biomass by 3.0 and 2.8 times, and increased leaf Bt protein content by 41.0 and 90.9%, and the ratio of N in Bt protein to total N by 9.3% and 15.6%, compared with the non-fertilization treatment, respectively; under moderate saline soil(total soluble salt content≈0.30%), N fertilization also improved plant biomass(1.4 times), total N content(98.8%) and Bt protein level(83.3%), but decreased the ratio of N in Bt protein to total N(7.3%). In the hydroponics experiment, plant biomass and the total N content in NO₃^--N treatment were higher than that in NH₄⁺-N with or without salinity stress, but the Bt protein levels was slightly lower than that in NH₄⁺-N treatment owing to decreased ratio of N in Bt protein to total N under salinity stress(11.0%). It is concluded that Bt protein concentration in leaves was influenced by nitrogen fertilization under salinity stress through increasing nitrogen uptake and/or altering the ratio of N in Bt protein to total N.
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  Effect of Planting Date on Yield and Yield Components of Two Different Cotton Genotypes

  LU He-Quan, LI Zhen-Huai, DONG He-Zhong, LI Wei-Jiang, TANG 　Wei, ZHANG Dong-Mei

  Cotton Science. 2012, 24(4): 312-317.

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  Selecting a suitable planting date is important for achieving high yield in cotton(Gossypium hirsutum L.), and is dependent upon cotton genotype. A two-year field experiment was conducted in Linqing, Shandong Province, to study the effects of cotton  genotype and planting date on yield, yield components, photosynthetic rate and harvest index. A split-plot design with three replicates was used for the study. The main plots comprised two cotton genotypes(the early-developing variety SCRC21 and late-developing cultivar SCRC28), while planting dates(early planting, mid planting and late planting) constituted the subplots. A significant interaction affecting economic yield was detected between genotype and planting date, mainly attributable to variation in boll numbers·m^-2 and lint percentage. For the early-developing cultivar SCRC21, plants sown in late April had the highest yield, 6.3% higher than those planted in mid-April; the yield was similar between plants sown in early and late April. For the late-developing cultivar SCRC28, plants sown in mid-April produced the highest cotton yield, which was 4.8% and 12% higher than for those planted in late April and early May, respectively. In the northwest of Shandong Province where film mulching is used, the early-developing variety is suitable for being delayed planted if no later than early May, and maximum yield can be obtained with being planted in late April. In terms of yield and economic index, the late-developing cultivar should be planted in mid-April, because later planting would significantly reduce cotton yield.
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  Effectiveness of the Micro-ditch and Overbite-membrane Planting Model for Cotton in a Dry and Alkaline Coastal Area

  WANG Zhan-Biao, LI Cun-Dong, LIU Yong-Ping, LIU Lian-Tao, SUN Hong-Chun, ZHANG Yong-Jiang, LI Hong-Qin, CHAI Wei-Dong

  Cotton Science. 2012, 24(4): 318-324.

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  Using the Cang 198 cultivar, with the traditional planting model as a control, we conducted field experiments to study the ecological and biological effects of the micro-ditch and overbite-membrane planting model(micro-ditch of 15 cm, plastic film mulching, sowing through holes perforated in the film) in Cangzhou saline fields. We studied soil volumetric water content and electrical conductivity, the percentages of seedling emergence and survival, the quality of the seedling environment and seedling physiological characteristics. The results indicated that soil volumetric water content under the micro-ditch and overbite-membrane planting model was significantly increased and soil electrical conductivity was significantly decreased compared to the traditional planting model. In the budding period, plant height, stem diameter and dry matter under the micro-ditch and overbite-membrane planting model were increased by 10.50%, 6.90%, 8.91%, respectively, compared to the traditional model, while SOD, POD activity and the proline content of functional leaves were reduced by 13.7%, 18.0%, and 11.7%, respectively,. Boll weight and yield increased by 7.34% and 8.07%. It can be concluded that the micro-ditch and overbite-membrane model collected rainwater and reduced salt, enhancing the quality of the seedling environment. Therefore the micro-ditch and overbite-membrane planting model can increase yield.
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  Comparative Studies on Physiological Characteristics of Main Stem Functional Leaves of Colored Cotton Varieties

  ZHANG Hai-Peng, ZHOU Qin, ZHOU Tao-Hua

  Cotton Science. 2012, 24(4): 325-330.

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  Use varieties of Wanmian 38(brown-fiber cotton), Wanmian 39(green-fiber cotton), and the parental material Wanmian 25 (white-cotton) to analyze the difference between the brown-fiber cotton and the green-fiber cotton in physiological characteristics of main stem functional leaves within two cotton varieties. The results showed that the contents of chlorophyll, soluble protein and soluble sugar in white cotton were the same as colored ones. The dynamics tend to peak at the full flowering stage. The three indices of colored cotton varieties were lower than the white cotton's at the whole stage, and rapidly decreased after the full flowering stage. The activities of SOD, POD and CAT within the two colored cotton varieties were quite similar at the whole development stage, with the activities of SOD, POD peaking at the full flowering stage and decreasing after the opening of bolls. Compared with brown-fiber cotton, the activities of SOD, POD, CAT of green-fiber cotton were much higher at the primary flowing stage, but much lower at the rest three periods. Specifically, MDA content in brown-fiber cotton was higher than those of green-fiber cotton and white cotton in initial stage of bloom. While in the boll opening stage, the MDA content of green-fiber cotton was higher. So that, when the green-fiber cotton was damaged under stress, activities of protective enzymes will be increased by functional leaves in order to minimize the damage to the cotton plant. While at the boll stage, activities of cell defense enzymes of green-fiber cotton decreased fast and resulted in senescence of cotton.
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  Absorption and Distribution of Foliar Applied Boron in Upland Cotton

  ZHANG Jun, WEI Chang-Zhou, LIANG Yuan-Hang, LI Mei-Ning, DONG Peng

  Cotton Science. 2012, 24(4): 331-335.

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  In this paper we studied the absorption and distribution of foliar applied boron in upland cotton(Gossypium hirsutum L.) in a hydroponic experiment. The results showed that both boron concentration and uptake in different cotton plant organs were the greatest in the leaves, followed by the stem, and lowest in the roots. boron absorption in the cotton plants increased significantly after foliar boron application except in roots. Compared with the control treatment, boron concentration in roots decreased by 41.9% when boron was partially applied on the leaves and by 11.2% when boron was applied to all leaves. Stalk boron concentration in the partial application treatment decreased by 5.9% compared to the control, but boron concentration in cotton leaves increased by 149.1%, significantly greater than the control treatment. In the partial application treatment, the amount of boron that was transferred to untreated leaves was only 5.2%, showing that boron absorbed by leaves can be partially transferred to conductivity organs but hardly enters other leaves. In the whole plant boron application treatment, the absorption of boron increased by 22.5% in the stalk and 203.7% in leaves.
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  Monitoring of Cotton Canopy Growth Status Based on Cotton Canopy APAR and FAPAR Data

  HUANG Chun-Yan, WANG Deng-Wei, CHENG Qi, ZHAO Peng-Ju, MA Qin-Jian-

  Cotton Science. 2012, 24(4): 336-340.

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  We cultivated two cotton cultivars in an experimental field, and recorded the photosythetically active radiation(PAR) of the cotton canopy using a linear quantum sensor at six key growth stages. The absorbed photosythetically active radiation(APAR) and fractional interception of absorbed photosythetically active radiation(FAPAR) were deduced from PAR data. The results showed that the highest APAR and FAPAR values were at the cotton flowering stage, the flowering to boll-forming stage, respectively. Their values decreased at the full boll stage and late boll stage, and reached a minimum at the boll opening stage. We used multivariate analyses for regression modeling between APAR/FAPAR, and leaf area index, ground cover, aboveground fresh biomass, and aboveground net primary production. The strongest relationship was between APAR and ground cover. Regression analyses showed that FAPAR and leaf area index were related via an exponential function. There were significant relationships between tested ground cover and estimated ground cover, and between tested LAI and estimated LAI, respectively. These results show that cotton APAR and FAPAR data can be used for real-time, nondestructive, and quantitative estimates of cotton canopy growth status parameters. These techniques will be useful for monitoring applications.
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  Correlation between Cotton Canopy CWSI and Photosynthesis Characteristics Based on Infrared Thermography

  CHENG Qi, HUANG Chun-Yan, WANG Deng-Wei, XIAO Li-Juan

  Cotton Science. 2012, 24(4): 341-347.

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   Using Fluke infrared thermal camera obtained canopy infrared thermal images at five main growth stages of cotton in an open experimental field including 2 cotton cultivars with 4 water treatments, meanwhile, use LI-6400 portable photosynthesis system test the net photosynthesis rate(P_n) , stomatal conductance(G_s), and use of LI-3100C area meter obtained leaf area index (LAI) ,respectively. Application of the image processing technology to acquire temperature of canopy sunlit leaves, and add wet artificial reference surface(WARS) temperature to empirical formulation defined by Jones, calculation of CWSI. Analysis of variation of CWSI against photosynthetic parameters P_n, Gs and LAI, cotton CWSI increased and P_n, G_s and LAI reduced correspondingly, the correlation between the average of CWSI and the average of Pn, Gs and LAI are negatively correlated (r_CWSI-Pn= -0.9182^**, r_CWSI-Gs=-0.8819^**, r_CWSI-LAI=-0.8661^**, n=16) under growth season, canopy CWSI and P_n, G_s and LAI can be synchronously reflecting cotton water stress status. The result showed that advanced infrared thermal image technology offered the potential to acquire spatial information on surface temperature in high resolution. It enables exclusion of background interference factors, calculation of CWSI is more accurate and it can also rapidly, effectively and precisely monitor cotton canopy water status.
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  Pathogenicity and ISSR Genetic Differentiation of Verticillium dahliae Isolates from Cotton Growing Areas of Hebei Province

  WANG Guo-Ning, ZHAO Gui-Yuan, YUE Xiao-Wei, LI Zhi-Kun, ZHANG 　Yan, ZHANG Gui-Yin, WU Li-Qiang, WANG Sheng-Fen, MA Zhi-Ying

  Cotton Science. 2012, 24(4): 348-357.

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  Fifty-two Verticillium dahliae isolates were recovered from 17 main cotton growing counties of Hebei Province. Culturing characteristics, pathogenicity and genetic variation based on ISSR marker were studied. According to culturing characteristics on PDA medium, the tested isolates could be classified into sclerotia type, filamentous type and intermediate type. Sclerotia type isolates which accounted for 51.9% were the most, followed by filamentous type (38.5%) and intermediate type (9.6%). In the growth chamber where the light, temperature and humidity were controlled automatically, the pathogenicity of 52 isolates was evaluated on seven cotton varieties with different disease resistance to Verticillium wilt. The results indicated that the isolates could be divided into three pathogenic types, i.e. strong (typeⅠ), mild (type Ⅱ) and weak (type Ⅲ), which accounted for 51.9%, 21.2% and 26.9% of the total, respectively. The nondefoliated isolate that had stronger pathogenicity than defoliated isolates was found. Generally, sclerotia- and intermediate- type isolates had strong or moderate pathogenicity, while filamentous type isolates had a widely pathogenic diversification. Cluster analysis based on disease index showed that 52 isolates had obvious pathogenicity differentiation that was not obviously related to their geographic origin. A total of 136 ISSR markers were amplified, and 80 markers that accounted for 58.8% of the total belonged to polymorphic markers. ISSR analysis indicated that the genetic variation of 52 isolates was small and related to geographic region.
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  Isolation, Identification and Activity of an Antagonistic Spore Bacterium against Cotton Verticillium Wilt

  ZHANG Dong-Dong, LI Shu-Na, GUO Xiao-Jun, ZHU Bao-Cheng

  Cotton Science. 2012, 24(4): 358-362.

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  A bacterial strain called Z-5 that had strong inhibition capability against Verticillium dahliae V-190 was obtained via preliminary and secondary screening with the improved agar plate diffusion method. Characterized by morphological and cultural features observation, physiological and biochemical experiments, and 16S rDNA sequence analysis, the Z-5 strain was initially identified as Bacillus malacitensis. Control of cotton Verticillium wilt by this antagonistic bacterium was assessed via potted plants. Results showed that the control efficacy of the antagonistic Z-5 strain on cotton Verticillium wilt was 76.05%. Preliminary results demonstrated that the antagonistic Z-5 bacterial strain could significantly inhibit the growth of V. dahliae.
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  Effects of Root Exudates and Phenolic Acids from Differently Resistant Cotton Cultivars on Verticillium dahliae

  ZHENG Qian, LI Jun-Hua, WEI Chang-Zhou, CHU Gui-Xin

  Cotton Science. 2012, 24(4): 363-369.

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  The root exudates of five cotton cultivars with different resistance to Verticillium dahliae were collected in Xinjiang, and their effects on V. dahliae pathogenic mycelium growth were studied to clarify the relationship between resistance and the root exudates. Results showed that the root exudates of resistant cultivars inhibited mycelium growth by 2%～40% compared with the control, the exudates of tolerant cultivars promoted mycelium growth early on but later inhibited it, and the exudates of susceptible cultivars promoted the mycelium. Phenolic acids in the root exudates of the five cotton cultivars were identified by high performance liquid chromatography(HPLC); gallic acid, chlorogenic acid, vanillic acid and p-coumaric acid were detected in different amounts in the different cultivars. Compared with the control, chlorogenic acid and p-coumaric acid inhibited mycelium growth at various concentrations; vanillic acid inhibited mycelium growth at higher concentrations, but at lower concentrations(< 400 mg·L^-1) promoted mycelium growth early on and inhibited it later; and gallic acid clearly promoted mycelium growth at concentrations of 0-800 mg·L^-1.
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  From Genomics to Proteomics: Great Molecular Progress in Cotton (Gossypium hirsutum L.)

  ZHOU Zhong-Hua, WANG 　Feng, CHEN Jin-Xiang

  Cotton Science. 2012, 24(4): 370-378.

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  Cotton genome project is being implemented and functional genomics has already played an essential role in cotton research. Great molecular progress has been made in genomic and proteomic field of cotton recently. This paper reviewed the progress of the cotton genomic and proteomics, particularly focusing on its genetic map, physical map, functional genome, proteomic techniques and cotton proteomics.