15 May 2010, Volume 22 Issue 3

    

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  Cloning of Stress-related Transcription Factors Gene from Cotton by Optimized TAIL-PCR

  LIANG Cheng-Zhen, ZHANG 　Rui, SUN Guo-Qing, MENG Zhi-Gang, ZHOU 　Tao, DING Zhong-Tao, GUO San-Dui

  Cotton Science. 2010, 22(3): 195-201.

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  ABF/AREB/ABI5/DPBF transcription factors belong to basic leucine zipper protein, this kind of transcription factors plays an important role in the signal transduction pathways of ABA-dependent abiotic stress. In this study, taking structure characteristics of ABF/AREB/ABI5/DPBF as the research object, we cloned three bZIP encoding genes by 3′ RACE successfully from cotton and also optimized the TAIL-PCR conditions. The further experiment results showed that we had got the integrated sequences of the ORF, 3′ UTR, as well as part of 5′ UTR sequence. The genes we cloned will provide candidate genes for resistance genetic engineering of cotton. Furthermore, the optimized methodology of TAIL-PCR will provide an easy and efficient technology for cloning of target genes.
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  The Optimization of ISSR-PCR System of Cotton by Uniform Design

  ZHU Yan-Fang, ZHU Shui-Jin, WANG 　Yang, LI Yong-Ping, MA Wen-Guang, HU 　Jin

  Cotton Science. 2010, 22(3): 202-208.

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  In present study，ISSR-PCR reaction conditions on Gossypium spp. were optimized. The factors which affect ISSR results of Gossypium spp. were investigated by uniform designs of two times. The results showed that 0.5 U Taq polymerase，40 ng template DNA，0.25 mmol·L^-1 dNTPs，0.5 μmol·L^-1 primer and 2.0 mmol·L^-1 Mg²⁺ in 20 μL ISSR reaction system might be the best combination. To validate the ISSR-PCR reaction system above，eight cultivars of Gossypium and three ISSR primers were employed in the research. It is indicated that the ISSR reaction system would be more productive and repeatable
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  Changes of Carbon and Nitrogen in the Subtending Leaf of Cotton Boll and Its Relationship to Biomass of Cotton Boll, Seed and Fiber

  ZHAO Xin-Hua, QU 　Lei, CHEN Bing-Lin, ZHOU Zhi-Guo

  Cotton Science. 2010, 22(3): 209-216.

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  Two cotton cultivars (KC-1, AC-33B) were used, with three nitrogen application rates (0, 240, 480 kg·hm^-2), standing for low, moderate and high nitrogen levels, respectively, in field experiments in Nanjing (118°50′E, 32°02′N, middle lower reaches of Yangtze River Valley) and Xuzhou (117°11′E, 34°15′N, Yellow River Valley), Jiangsu Province, which stand for the different ecological conditions, to investigate the relationship between the changes of carbon and nitrogen in the subtending leaf of cotton boll and biomass of cotton boll. The results showed that the changes of total carbon and total nitrogen content in subtending leaf of cotton boll could be simulated with quadratic: Y=at²+bt+c (Y stands for the total carbon or total nitrogen content (%), t stands for boll age (d)). And the changes of C/N ratio could be simulated with Logistic equation. Correlation analysis indicated: (1) The average rate of cotton boll, seed and fiber biomass during the speedy accumulation period could be improved, the duration of cotton boll and fiber biomass could be shortened and the duration of the seed biomass speedy accumulation could be prolonged when the date of reaching the minimum content of the total carbon was earlier and the minimum content of the total carbon was higher. (2) The period of cotton boll and fiber biomass speedy accumulation could be prolonged as the date of reaching the minimum content of the total nitrogen was later. And the minimum content of total nitrogen was higher, which resulted in prolonging the cotton seed biomass speedy accumulation period. (3) The duration for cotton boll and fiber biomass speedy accumulation could be prolonged by lengthening the speedy increasing duration and slowing the average rate of C/N ratio increase during the speedy increasing period. Therefore, the changes of carbon and nitrogen in the subtending leaf of cotton boll is tightly related to the cotton boll biomass, boll development could be improved by adjusting the content of carbon and nitrogen and the C/N ratio in the subtending leaf.
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  Somatic Embryogenesis and Plant Regeneration in Wild Cotton Species of G.tomentosum

  ZHANG 　Qiao, WANG Jing-儿, YE Chun-Li, YAN Shu-Feng, CHEN Jin-Hong, HONG Cai-Xia, ZHU Shui-Jin

  Cotton Science. 2010, 22(3): 217-223.

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   The wild cotton species of G.tomentosum has some outstanding characters such as the higher resistance to pests and tolerance to drought. However, the upland cotton germplasm with the trait has not been obtained through sexual hybridization between G.hirsutum and G.tomentosum due to the distant relationship between the wild species and upland cotton. Protoplast fusion technique which depends on the success in the somatic culture of two different species can avoid sexual incompatibility. The callus inducing and somatic embryo germination of G.tomentosum were studied through the regulation of culture conditions such as plant growth regulators(PGRs), PEG, CuSO₄ and AgNO₃. The results indicated that under the same condition the frequency of somatic embryo germination and the plant regeneration ability of G.tomentosum were lower and the plant regeneration was more difficult than Coker 201. But through the regulation of culture conditions, we can get many embryoids and few of them converted to plantlets. The callus of G.tomentosum induced in the medium with the hormone combination of 0.1  mg·L^-1 KT + 0.1 mg·L^-1 2, 4-D held good for cell differentiation, and it was better for proliferation of the embryogenic callus in the medium with 0.5 mg·L^-1 IBA + 0.2 mg·L^-1 KT. The medium supplied with PEG was not helpful for initiation of somatic embryos. But the medium of MSB₅ with 3.5 mg·L^-1 CuSO₄+0.5 mg·L^-1 IBA + 0.2 mg·L^-1 KT, or 3 mg·L^-1 AgNO₃+0.5  mg·L^-1 IBA+ 0.2 mg·L^-1 KT were good for the proliferation of embryogenic callus and germination of somatic embryo. This report provides an useful technique for somatic culture of the wild cotton species, and also ensures the protoplast fusion feasibly between these wild cotton species and other cotton species to centralize several good traits into a new germplasm. However, it is needed to study further about the various culture factors and conditions for building a quick and high efficient regeneration technique of G.tomentosum.
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  nteraction of Plant Density with Retention of Vegetative Branches on Yield and Earliness of Cotton (Gossypium hirsutum L.)

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

  Cotton Science. 2010, 22(3): 224-230.

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  Optimum plant density and retention of vegetative branches(VB) have been widely studied as a single practice in cotton production, but their combination to approach better yields and benefits was less studied. Since effective combination of plant density and VB retention may further reduce cost and increase yield, it is very important to conduct profound studies on interaction of plant density and VB retention. Using upland cotton Lumianyan 28, a multi-site field experiment was conducted in Linqing City, Xiajin County and Huimin County of Shandong Province respectively, to study interaction of plant density and VB retention on yield, yield components, earliness, and economic index in 2008. A split-plot design with four replications was used for the study. The main plot was plant pruning(removal and retention of VB), while plant density(3.00, 5.25, 7.50 and 9.75 plants·m^-2) constituted the subplots. Significant interaction was detected between plant density and plant pruning. Plants without VB produced the lowest and highest cotton yield at 3.00 plant·m^-2 and from 5.25 to 7.50 plants·m^-2 respectively, while those with VB produced the highest and lowest cotton yield from 3.00 to 5.25 plants·m^-2 and from 7.50 to 9.75 plants·m^-2 respectively. Under VB removal cotton yield at 5.25 plants·m^-2 was 9.7% higher than at 3.00 plants·m^-2, but under VB retention cotton yield at 3.00 plants·m^-2 was similar to that at 5.25 plants·m^-2 and 15.3% higher than that at 9.75 plants·m^-2. Both plant pruning and plant density significantly affected yield components. Boll weight was higher under VB removal than retention, and the number of boll per m2 increased as plant density raised. There existed significant interaction among site, plant density and plant pruning. Earliness under VB removal was higher than under VB retention, while earliness at 5.25~7.50 plants· m^-2 was better than at 3.00 and 9.75 plants·m^-2. No interaction of plant density and plant pruning on ratio of seedcotton to stalk, but VB retention and elevated plant density significantly increased biological yield and decreased ratio of seedcotton to stalk. Plant density and plant pruning affected biological yield, economic index and yield components individually or interactively, thus resulting in a significant interaction on economic yield. VB retention can compensate for yield loss due to lower population under low plant density，while VB removal is still beneficial to yield increase under middle and high plant density in Shandong Province.
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  The Effects of Drought on Endogenous Hormone Contents and Balance in Main Stem Leaves of Cotton

  LI Dong-Xiao, LI Cun-Dong, SUN Chuan-Fan, SUN Hong-Chun, LIU Lian-Tao, ZHANG Yong-Jiang, XIAO 　Kai

  Cotton Science. 2010, 22(3): 231-235.

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  As Lumianyan 28 to be the material, the dynamic changes of endogenous hormone contents in main stem leaves of cotton were studied by enzyme-linked immunosorbent assay, two different water treatments drought (the earth RWC<60%) and CK (the earth RWC as normal) being set. The results indicated that ZR contents in main stem leaves both showed single peak curves under two treatments, and were very significantly higher but later under drought condition than that of CK. The GA₃ content of drought leaves showed second-order peak value, which indicated plants entered the reproductive stage earlier. IAA content increased at metaphase in drought stress, but there was no obvious difference in variation tendency between two treatments. It took on a higher peak value of ABA in early drought leaves, and later, ABA ascended slowly to promote premature. Also, drought influenced the dynamic balance of ZR/ABA、GA₃/ABA、IAA/ABA and (ZR+GA₃)/ABA. It took longer actions of ZR under drought condition; GA₃/ABA was closely related in the hormone mutual actions; IAA/ABA played an important physiology role at later growth even in water deficiency.
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  Comparison of Relative Enzymes Activities during Fiber Development Period between Brown and White Fiber Cotton Cultivars

  CHEN Yun-Feng, WANG Ya-Nan, LIN 　Yi, CAI Yong-Ping, LI Ting-Chun, LI Zheng-Peng, FAN Hong-Hong

  Cotton Science. 2010, 22(3): 236-241.

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  This experiment was carried out to study the effect of relative enzymes activities on brown cotton fiber qualities, three kinds of brown cottons (dark brown, mid-brown and light brown) were used to detect the dynamics of relative enzymes activities in contrast with one white cotton. The results showed that the length of fiber, lint weight and lint percentage of brown cotton were lower than white cotton. During the fiber development, the elongation rate of brown cotton was lower and the elongation term was shorter than white cotton. Comparing with white cotton, brown cotton had shorter term, lower cellulose accumulation rate and cellulose content for fiber. At the early periods of fiber development, SS activity was lower in brown fiber, while POD and IAAO activities were higher. At the later periods of fiber development, β-1, 3-glucanase and POD activities were lower in brown fiber.
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  Differences in SOD, POD, CAT Activities and MDA Content and their Responses to High Temperature Stress at Peak Flowering Stage in Cotton Lines with Different Tolerance to High Temperature

  DENG Jiang-Ming, XIONG Ge-Sheng, YUAN Xiao-Ling, JIA 　Fei, LIU 　Zhi-

  Cotton Science. 2010, 22(3): 242-247.

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  Activities of SOD, POD, CAT protective enzymes and MDA content in main-stem leaf and anther at different developmental stages and their responses to high temperature stress at peak flowering stage were analyzed comparatively using three cotton lines exhibiting tolerance, moderate sensitivity and sensitivity to high temperature in the present paper. The protective enzyme activities in main-stem leaves were nearly same among the three cotton lines at seedling and peak squaring stages, but significantly higher in tolerant line than susceptible ones at peak flowering and boll-setting stages. The MDA content was markedly lower in tolerant line than susceptible ones from peak squaring stage. Activities of protective enzymes in cotton anther increased with developmental processing, and remarkably higher SOD and POD activities in anther at the pollen maturing stage were measured in tolerant line. During the whole developmental period of anther, CAT activity was greater, while the MDA content was lower distinctly in tolerant line than susceptible ones. The protective enzyme activities were inhibited, and the MDA content increased excessively in main-stem leaves and anther of the three cotton lines under high temperature stress at the peak flowering stage. However, changes of decreasing in activities of protective enzymes and increasing in the MDA content in leaves and anther were less obviously in tolerant line than susceptible ones.
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  Response of Plant Height to the Exogenous Hormones of Dwarf Cotton (Gossypium hirsutum L.)  Lu-ai No.1

  LIU Ai-Yu, CHEN Jin-Xiang, LI Rui-Lian, YI Jiu-Hong, FENG Zheng-Rui

  Cotton Science. 2010, 22(3): 248-253.

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  The study was undertaken to assess the sensitivity of plant height of Dwarf cotton (Gossypium hirsutum L.)  Lu-ai No.1 to ectohormone through spraying different concentrations of hormones at the budding stage and flower-boll stage. The results showed that: Lu-ai No.1 was more sensitive to the exogenous hormones than the CK, various concentrations of GA₃ can promote  plant height of Lu-ai No.1 significantly, especially under 100 mg·L^-1 and 150 mg·L^-1 concentration, the promoting rate of 100.70% and 59.15% for Lu-ai No.1, while 27.51% and 27.88% for TM^-1( CK), respectively. ABA significantly inhibited the plant height of Lu-ai No.1, the higher concentration, the stronger inhibition, the promotion rate was -31.03% under 40 mg·L^-1. Lower concentration (20 mg·L^-1) of the IAA treatment can significantly promote the plant height of dwarf Lu-ai No.1 and reverse under higher concentrations; higher concentration of KT treatment significantly inhibited the plant height of Lu-ai No.1, but lower concentrations indistinctively promoted. Mixed spraying with four kinds of hormones showed up antagonistic effect on promoting plant height of Lu-ai No.1, with much lower promoting rate than that of GA₃ treatment alone.
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  Comparison of Low Temperature-induced Protein in Cotton Seedlings with Different Chilling Resistance

  LI 　Rui, LI Sheng-Quan, FAN Yue-Xian

  Cotton Science. 2010, 22(3): 254-259.

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  By using two-dimensional electrophoresis and mass spectrometry techniques, the proteomes in different chilling-resistant levels cotton Jin3047 (It has obvious chilling resistance) and 410 (It has less chilling resistance) before and after low temperature treatments were comparatively analyzed. Results showed that the proteomes had a significant difference in different chilling-resistant level cotton varieties, within the range of pH4~7, there was a significant difference in the expression level of 50 protein spots in the protein profiles of cotton seedlings Jin3047 before and after low temperature treatment, and 30 of 50 protein spots were expressed. After low temperature treatment, the abundance value of 19 protein spots increased, with the absolute abundance being higher in Jin3047 than in 410, while that of 11 protein spots decreased, with the absolute abundance being lower in Jin3047 than in 410. The expression level of 8 protein spots in Jin3047 was characterized and the expression quantity was increasing, but no expression in 410. The differential protein spots were detected by the mass spectrometry, and identified by compared with the retrieval database. A total of 30 protein spots fingerprints were obtained, among which, 20% were of stress protein. These variously expressed proteins might play an important role in the chilling resistance of Jin3047.
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  The Effect of Different Density to Canopy-Microclimate and Quality of Cotton

  LOU Shan-Wei, ZHAO 　Qiang, GAO Yun-Guang, GUO Ren-Song, A Bu-Li-Ke-Mu, ZHANG Ju-Song

  Cotton Science. 2010, 22(3): 260-266.

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  In order to study the effects of different density to the micro-climate of cotton canopy and the relationship between cotton physiology characteristic and its quality, we has designed five kinds of different densities in field study. The results indicated that the transmittance of different crown level after first flowering dropped rapidly, in different crown level the transmittance A3 (180000 plants per hectare) became the highest. In the flowering season air temperature was the highest by A3 when the crown level was 20 cm. After the boll opening, the regularity was not obvious. In the flowering season relative humidity was high, but its change was small. The relative humidity was the stablest by A3, and the change was the smallest. The change was big by other treatments. In the boll opening period, change tendency of relative humidity was consistent basicly in different crown levels, the relative humidity was a little higher in the morning and evening, the regularity was not obvious among different treatments. The yield of ginned cotton in A3 was the highest, the LAI peak appeared during the cotton bolling period, it still maintained 2.5~2.6 at the boll opening stage. The leaves of tenth fruit branches senesced in August 14th, after 5 days began the eighth fruit branches, A3 could maintain the normal physiological condition longer. Cotton textile fiber quality of different treatments had no significant difference, the increasing of the density could promote the fiber quality, but it had negative effect on the fiber quality beyond the density of A4(225000 plants per hectare).
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  Analysis on the Relationship between Heterosis of Quality Traits Fiber Colors

  SUN Jun-Ling, HUO Shu-Ming, ZHOU Zhong-Li, JIA Yin-Hua, PAN Zhao-E, PANG Bao-Yin, DU Xiong-Ming

  Cotton Science. 2010, 22(3): 267-272.

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  Seven kinds of hybrid combinations were made based on four kinds of color fiber lines including white, green, brown, dark brown. The differences of fiber color and the fiber quality traits, the correlation between fiber color and the heterosis of the fiber quality traits were analyzed. The results showed that: the average super-parent heterosis between the cross of the dark brown and green line was strongest. The average mid-parent heterosis in the combinations between the white and dark brown, dark brown and green lines were better. The correlation coefficients between super-parent heterosis and their parents  in absolute value of fiber length, strength, micronaire traits were very significantly different. But the correlation between mid-parent heterosis and their parents were not different in a significant level. The correlation coefficients between the fiber color and fiber length, strength traits, super-parent and mid-parent heterosis were -0.3601 and -0.4487, -0.3978 and -0.3895, respectively, all were very significant. The correlation coefficient of fiber color and the F₁ fiber length was -0.6045, significant at 0.01 levels. In breeding, the heterosis can be used to improve fiber length and strength simultaneously. The crosses of dark brown and green, green and green, brown and green combinations can be used for improving fiber quality with dark color fiber. The darker the fiber colors of two parents were, the weaker the super-parent and mid-parent heterosis of fiber length and strength traits were. So it is difficult to improve the fiber quality of color cotton.
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  Progresses on QTL Mapping for Cotton Resistant to Verticillium Wilt

  FENG Chang-Hui, ZHANG Sheng-Xi, SHI Ren-Hui, FU Li-Li, BAI Jing, CHEN Yin-Hua

  Cotton Science. 2010, 22(3): 273-278.

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  The development of QTL mapping make it easy to dissect the genetic basis of quantitative traits into Mendelian factors, and the genetic basis of quantitative traits could be better understood. So the technique could lay a foundation for the acceleration of improvement on cotton resistant to Verticillium wilt through molecular breeding.In the light of problems as encountered in QTL mapping for cotton resistant to Verticillium wilt, we made a detailed review that focused on the theory,progresses on QTL mapping for cotton resistant to Verticillium wilt. The application of QTL mapping for cotton resistant to Verticillium wilt to genetic improvement in the future was discussed also in this paper.
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  Research Progress of Premature Senescence in Cotton

  ZHANG Hai-Na, ZHANG Xiang-Yun, LI Jun-Lan, CUI Shu-Fang, JIN Wei-Ping, WANG Guang-En

  Cotton Science. 2010, 22(3): 279-284.

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   It is a very urgent research task as the cotton premature senescence is more and more serious. It is only make clear the causes and mechanisms which induced the cotton premature senescence that we can make the reasonable prevention measures. So lots of researchers have researched cotton premature senescence in different aspects. They discussed the cotton premature senescence affected by stress surrounding, soil, planting measure and so on, analyzed biochemical reaction change in premature senescence cotton, tried to breed new varieties with molecular biology measure. This article summarized some important viewpoints about cotton premature senescence shape characters, formation causes, physiochemical reaction, and advised many reasonable and effective cultivation techniques in preventing premature senescence. Planting varieties resistant to premature senescence is the most effective measure. We should do further research to prevent drastically cotton premature senescence.