15 January 2013, Volume 25 Issue 1

    

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  Cloning of Aquaporin Gene GhNIP6.1 from the Upland Cotton and Its Expression in Different Cotton Tissues

  GONG Yuan-Yong, GUO Shu-Qiao, SHU Hong-Mei, NI Wan-Chao

  Cotton Science. 2013, 25(1): 1-8. https://doi.org/10.11963/cs130101

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  Primers were designed according to the assembled ESTs of Gossypium hirsutum, and an aquaporin gene GhNIP6.1 was obtained from upland cotton Sumian 18 in this experiment by RT-PCR. The gene contains an ORF comprising of 903 nucleotides which encodes 300 amino acids with a predicted molecular mass of 30.97 kD and theoretical isoelectric point of 8.99．Bioinformatics analysis indicates the GhNIP6.1 exhibited a typical structure with six trans-membrane domains which linked by five loops, loop A, C and E were at outside of membrane, loop B, D, N terminal and C terminal were at inside of membrane. N terminal has 79 amino acids and C terminal has 18 amino acids. The amino acid sequence of GhNIP6.1 has the conserved peptides of MIP family. The result of predicted protein 3D structure of GhNIP6.1 showed that it was highly similitude with that of AtNIP6.1. The subcellular localization of GhNIP6.1 maybe localize at plasma membrane, which is consistent with AtNIP6.1. Phylogenic analysis displays GhNIP6.1 has the highest similarity with Arabidopsis thaliana AtNIP6.1 protein. A genomic DNA sequence with 2021 bp clone containing an aquaporin was isolated from the upland cotton in this experiment, and the GhNIP6.1 gene contains eight introns and seven exons. All exon／intron junctions of the gene contain the gt／ag consensus splicing site. Semi-quantitative analysis of GhNIP6.1 expression showed that the gene expressed extensively in cotton roots, stems, leaves, except cotyledon, and leaf is the most highly expressed tissue.
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  Studies on Construction Method of Cotton Bacterial Artificial Chromosome Library

  GAO Hai-Yan, WANG Sheng-Fen, LIU Fang, PENG Ren-Hai, ZHANG Yan, MA Zhi-Ying, WANG Kun-Bo

  Cotton Science. 2013, 25(1): 9-16. https://doi.org/10.11963/cs130102

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  Bacterial artificial chromosome (BAC) library is an important genome resources to such research as genome sequencing, map-based cloning, molecular markers, and physical mapping. On the base of the construction of BAC library for Gossypium herbaceum var. africanum, this paper presents an exhaustive analysis on details and notices of the BAC library construction process. It includes extraction of high molecular weight (HMW) nuclear DNA, determination of the optimized enzyme for partial digestion of HMW DNA, two rounds of size fractionation, recovery of large fragments DNA, ligation and transformation of large fragments of DNA and storage of BAC library. Thus being able to supply an experience for constructing high efficiency cotton BAC library.
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  Effect of Medium, Ovule Age, and Hormone Combinations on the Fiber Growth and Development of Cotton Ovule Culture in vitro

  YE Chun-Yan, OU Ting, CHEN Jin-Hong, HE Qiu-Ling, ZHU Shui-Jin

  Cotton Science. 2013, 25(1): 17-23. https://doi.org/10.11963/cs130103

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  In order to establish a suitable system for fiber growth and development during cotton ovule culture in vitro, effects of medium types, hormones and hormone combinations, ovule age etc. were studied, using CCRI 49 as material. As well, based on the established experiment system for ovule culture in vitro, the genotype effect in ovule culture was studied. The results showed that BT basic medium is the best medium for cotton ovule culture in vitro. Liquid culture was better than solid one although both of them could induce the fiber growth and development during the ovule culture. All the ovules sampled before flowering and after flowering can be used as explants in ovule culture, but the best one for fiber growth and development was the ovules sampled at flowering day. Hormone was necessary for fiber growth and development during ovule culture in vitro, all the hormones used in the experiment such as GA₃, IAA, NAA, Ethylene, and BR could induce the fiber growth and development, and GA₃ was best one. The effects of hormone combinations on fiber growth during the ovule culture in vitro were better than that with single hormone individually, and 5.0 μmol·L^-1 IAA+5.0 μmol·L^-1 GA₃ was the best one among the hormone combinations used in the experiment. The fiber growth and development during the cotton ovule culture in vitro was different significantly among the genotypes used in the experiment, and the best hormone or hormone combination might be different for the different genotypes.
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  QTL Mapping of Fiber Quality Traits for Elite Hybrid Derived-RILs on Chromosome 22sh in Cotton

  YAO Jin-Bo, CHEN Wei, ZHANG Yong-Shan, GUO Xiang-Mo, CHU Li, LIU Hai-Ju

  Cotton Science. 2013, 25(1): 24-29. https://doi.org/10.11963/cs130104

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  A population of 104 recombinant inbred lines (RILs) was developed by single seed descended from the cross of upland cotton(Gossypium hirsutum L.) TM-1 and its substitution line, CSB22sh. Agenetic linkage map consisting of 61 loci and covering 76.93 cM was constructed using SSR markers, with the average distance of 1.26 cM between two markers. Composite interval mapping was used to identify the quantitative trait loci (QTL) in RILs, 12 QTLs for fiber quality were independently detected in four environments, whereas 4 QTLs were found in joint analysis. These QTLs explained 2.45%~21.11% of the phenotypic variance. There were 9, 1, 1 and 1 QTLs simultaneously significant in one, two, three and four environments, respectively. There were two fiber length and two fiber uniformity QTLs in joint analysis, which explained 14.37% ~ 19.97% of the phenotypic variance and in same position respectively. Notably, a stable fiber length QTL qFl-22-2 was detected both in joint analysis and in four environments of separate analysis, which might be of special value for marker-assisted selection.
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  Responses of Growth in the Seedling Period of Transplanted Cotton to the Shading Treatment

  LI Fei, MAO Shu-Chun, HAN Ying-Chun, FAN Zheng-Yi, WANG Guo-Ping, LI Xiao-Xin, LIU Fang

  Cotton Science. 2013, 25(1): 30-36. https://doi.org/10.11963/cs130105

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  This study was to investigate changes of substrate seedling transplanted cotton in seedling period under different shading treatments. The results showed that: during the recovering stage, the soluble sugar content and starch accumulation capacity of different shading treatments has been great influenced significantly. After recovering-stage, the change trend of soluble sugar content and starch accumulation capacity was broadly the same, but there were certain differences in different treatments. SOD activity and POD activity were all going up during the recovering-stage, and then decreasing to a steady level. The SOD activity and POD activity of shading groups were lower than the control treatment. After recovering-stage, the leaf area, fresh weight, dry weight and plant height of shading groups were larger than those of the control treatment. The degree of shading during seedling period can alleviate the relative adversity that the substrate seeding-raising transplanted cotton located and be beneficial to the growth and development of cotton seedlings.
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  Effect of Field Arrangement in Apricot-cotton Intercropping on Cotton Plant Growth and Tempo-spatial Distribution of Boll

  LIU Yu-Na, CHEN Fei-Hu, XU Xin-Qiang, ZHANG Ju-Song, XU Wen-Xiu, ZHANG Yong-Qiang

  Cotton Science. 2013, 25(1): 37-44. https://doi.org/10.11963/cs130106

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  In order to find out high-yielding cotton cultivation of agroecology in apricot-cotton intercropping which planting spacing is 3 m×6.5 m, and height is 5.45 m, the experiment is conducted with three field arrangements on cotton plant growth and tempo-spatial distribution of boll. The results showed that field arrangement with 10 cm plant spacing, 10 rows (B) has the lowest length-width ratio, and a better performance in main stem leaf number, width of the top fourth leaf and fruit branch number than others. Stem diameter was positively related to plant spacing. Cotton individual of dry matter was higher than others, dry matter of gross was positively related to individual; dry matter of net was positively related to density. Shade delayed the growth of cotton,leading to three treatments were the most of summer bolls and autumn bolls; bolls mainly distributed close to stem in horizontal, as plant spacing increased; longitudinal were main compose of middle bolls and lower bolls. Lint yield of gross plant in plant spacing of 10 cm (B) was the highest, higher 4.69% and 19.96% than plant spacing of 12 cm and 8 cm; Lint yield of net plant in plant spacing of 8 cm was the highest, higher 7.18% and 30.91% than plant spacing of 10 cm and 12 cm.
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  Determination of Bt Protein in Transgenic Cotton Seeds by Double Antibody Sandwich Enzyme-linked Immunosorbent Assay

  SUN Shuo, ZHANG Kun, TAN Gui-Yu, CAO Zhen, NAN Tie-Gui , WANG Bao-Min

  Cotton Science. 2013, 25(1): 45-50. https://doi.org/10.11963/cs130107

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  Bt Cry1Ac protein sandwich enzyme-linked immunosorbent assay was developed with anti-Bt Cry1Ac protein mouse monoclonal antibody (mAb) and rabbit polyclonal antibody produced by Chemical Regulation Research Center (CRRC) of China Agricultural University (CAU). The linear range of the method was 0.78-50.0 ng·mL^-1. The linear equation was y = 0.6634x-1.7387, and the determinative coefficient was 0.992. The specificity obtained with the established assay was confirmed and verified by the commercial Bt Cry1Ac protein kit and could be qualitatively and quantitatively used to detect the Bt Cry1Ac protein. By the assay, identification of hybrid F₁ and F₂ with either of the parents is non-transgenic Bt cotton showed that, all the seeds of F₁ were positive and the ratio of seeds positive to negative of F₂ was 3∶1. The assay was also used to imitate AP detection and the limit of AP was 1∶110 for single seed which contained more than 140 ng of Bt Cry1Ac protein.
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  Analysis on the Changes of Physiological Indices of Young Leaves Infected by Lygus lucorum of Transgenic Bt Cotton

  LIU Yu, ZHANG Pei-Tong, CHEN Bing-Lin, GUO Wen-Qi, ZHANG Heng-You

  Cotton Science. 2013, 25(1): 51-56. https://doi.org/10.11963/cs130108

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  The experiment was carried out in 2011, at the Institute of Industrial Crops, Jiangsu Academy of Agricultural Sciences. The Bt transgenic cotton cultivars(Sukang 310) and its recurrent parent(Simian 3) were used as the experimental materials to investigate the changes on physiological indices of the young leaves after the cotton were infected by Lygus lucorum. Infected by Lygus lucorum, the free amino acids contents, the soluble protein contents and the chlorophy contents of the young leaves of both the transgenic Bt cotton and non-transgenic Bt cotton were decreased significantly, and the condensed tannin contents and phenylalanine ammonia-lyase(PLA) activities of young leaves were increased significantly, in both bud stage and boll stage. Infected by Lygus lucorum, the soluble sugar content of young leaves of transgenic Bt cotton was increased, while that of non-transgenic Bt cotton was decreased. It's indicated that after damaged by Lygus lucorum, the nitrogenous metabolism of young leaves was decreased, and led to the chlorophyll content decrease. It's harmful to the photosynthesis of the leaves grown mature. Increasing in the condensed tannin contents and PLA activities was advantageous to withstanding infecting of Lygus lucorum. While the condensed tannin contents and PLA activities of the young leaves damaged by Lygus lucorum of the transgenic Bt cotton was lower than that of the normal young leaves of non-transgenic Bt cotton. It was one of the possible cause of the change on the population dynamics and damage of Lygus lucorum. Increasing in the soluble sugar content was advantageous to the infecting of Lygus lucorum. While damaged by Lygus lucorum, the increasing in soluble sugar content of young leaves of transgenic Bt cotton was advantageous to infecting of Lygus lucorum. It was another possible cause of the change on the population dynamics and damage of Lygus lucorum.
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  Sampling Estimation of Suitable Quantity of Test Sites in Cotton Variety Regional Trials

  XU Nai-Yin, JIN Shi-Qiao

  Cotton Science. 2013, 25(1): 57-62. https://doi.org/10.11963/cs130109

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  The sampling populations of four check varieties, namely, Simian 3, Xiangzamian 2, Xiangzamian 8 and Ezamian 10 were constructed and analyzed based on their performances in earliness, yield, fiber quality and agronomic traits in 84 to 270 time tests in national cotton regional trials in the Yangtze River Valley during 2000－2011 period. And the adequate and necessary quantity of regional trial sites to identify cotton main characteristics were estimated using random sampling method within the cultivar population in accordance with the responding sampling estimation accuracy ratio, so as to investigate the theory and methodology in determining the suitable quantity of cotton regional trial locations in the Yangtze River Valley, and also act as a useful demonstration for other cotton planting regions and even other crops in regional trial sites determination. Research results showed that the number of sites demanded was closely related with the target selection trait and its variation degree, so the lint yield with the highest degree of variation demanded most quantity of trial locations under the same estimation precision level. At present, the estimation accuracy of lint yield reached 90% in national cotton regional trials with eighteen trial sites in the Yangtze River. In case the estimation accuracy was set to 93%, additional sixteen test sites must be arranged, and the subsequent increase of test sites was estimated to be less effective in estimation accuracy enhancement.
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  Effect of Combination Stress of Temperature and Humidity on Insecticidal Protein Expression of Bt Transgenic Cotton

  WANG Yong-Hui, CHEN Jian-Ping, CAI Li-Wang, ZHANG Xiang, CHEN Yuan, CHEN De-Hua

  Cotton Science. 2013, 25(1): 63-67. https://doi.org/10.11963/cs130110

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  Using insect-resistant transgenic cotton Sikang 3 as material, four combined stress was desigened to study the effect of combination stress of temperature and humidity and its recovery on insecticidal protein expression of Bt transgenic cotton which was high temperature-humidity, low temperature-humidity, high temperature with low humidity and low temperature with high humidity. The results showed that combined stress decreased insecticidal protein content. Under the same growth period, the decrease amplitude of Bt content was lowest with high temperature-humidity stress, while highest with low temperature-humidity stress. However, the recovery level of Bt content after released from combined stress was highest with high temperature-humidity stress, while lowest with low temperature-humidity stress. Under the same stress, the decrease amplitude of Bt content was low at peak square period, middle at peak flowering period, high at peak boll period under four combined stress, however, the recovery level of Bt content after stess stopped performed reverse trend. These results suggested that the decrease amplitude of Bt content under combination stress of  temperature and humidity stress and recovery level of Bt content after stress stopped was closely associated with stress types and cotton development stages.
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  Effects of Transgenic Cry1Ac plus Cry2Ab Cotton on Pest Survival，Growth and Enzymes Activity of Three Kinds of Pests

  LUO Jun-Yu, CUI Jin-Jie, ZHANG Shuai, XIN Hui-Jiang, WANG Chun-Yi

  Cotton Science. 2013, 25(1): 68-72. https://doi.org/10.11963/cs130111

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  Using the transgenic Cry1Ac cotton and non-transgenic cotton as control, the effects of transgenic Cry1Ac plus Cry2Ab cotton on survival, growth and enzymes activity of three types of pests (Helicoverpa armigera Hübner, Spodoptera liture and Spodoptera exigua Hübner ) were studied. The results show that, the difference of chronic toxicity to three types of pests fed with leaves from different cotton varieties is significant, and resistance of every type cotton varieties to these pests is different too. Compared to the conventional variety CCRI 49, the growth speed of three pests fed with transgenic Cry1Ac+Cry2Ab gene cotton and transgenic Cry1Ac cotton is slower, and the weight is lighter. Meanwhile enzymes activity change with different degrees too, but the changes of each enzyme activity have some difference.
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  Effects of Water and Fertilizer Regulation Strategies on Cotton Canopy Structure and Yield under Mulch Drip Irrigation

  YAO Qing-Qing, YANG Tao, MA Xing-Wang, XUE Xiang-Rong, NIU Xin-Xiang, WANG Bin

  Cotton Science. 2013, 25(1): 73-80. https://doi.org/10.11963/cs130112

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  Determined cotton leaf area index and other canopy characteristics in full growing period under mulch drip irrigation of the conventional irrigation fertilization, optimized cotton irrigation, optimized fertilization, and analyzed the dynamics of cotton canopy structure and its relationship with yield. The results showed that: conventional fertilization with N and P in the conventional irrigation could make the cotton LAI (Leaf area index) value and MFIA (Mean foliage inclination angle) value significantly increase, TC (Transmission coefficient for diffuse penetration) value and T (Transmission coefficient for radiation penetration) value significantly reduced, which can make the number of bolls per unit area and boll weight significantly increase, and then led to the seed cotton yield significantly increased; N, P, K balanced fertilizer can make the cotton LAI value and MFIA value slightly decreased, reduce TC value and T value, which can increase boll number and then improve seed cotton yield. In conventional fertilization, reducing water amount, optimizing irrigation frequency and irrigation water of the growth period can increase the cotton LAI value and MFIA value, reduce TC value and T value, which can increase boll weight and then make seed cotton yield significantly increased.
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  Discrimination on the Relationship among Cotton Premature Senescence, Red Leaf Disease and Alternaria Leaf Spot Disease

  QI Fang-Jun, JIAN Gui-Liang, LI Jia-Sheng

  Cotton Science. 2013, 25(1): 81-85. https://doi.org/10.11963/cs130113

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  Relationships among cotton premature senescence, cotton red leaf disease and Alternaria leaf spot disease were comprehensively analyzed. Based on the different causes, cotton premature senescence could be divided into three types, which were physiological, pathological premature senescence and compound premature senescence mixed with physiological and pathological factors. Among them，the often appearance of compound premature senescence always caused serious harm for cotton production in recent years. The causes of compound premature senescence often happened at mid to late cotton growth period was particularly explained as that cotton leaves enter into aging process, when cotton encounter adverse environmental conditions which can cause physiological impairment and decrease in disease resistance to cotton leaves, and make the cotton leaves being infected by Alternaria leaf spot pathogens Alternaria alternata. In addition, a comprehensive prevention technology for cotton premature senescence was put forward on the basis of our investigation results in recent years.
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  Application Study about OTSU Optimal Threshold Method in Cotton Seedling Recognition

  LIU Guang-Rui, HUANG Zhen, MAO Shu-Chun, WANG Guo-Ping, BI Jing-Kai

  Cotton Science. 2013, 25(1): 86-89. https://doi.org/10.11963/cs130114

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  Cotton seedling recognition and depart is the key technology to realize transplanting automation, using visual picture processing technology, the cotton seedling recognition is studied. To the cotton seedling picture got by vision, based on the color analysis of cotton seedling and its background, the color pictures of cotton seedling expressed at RGB color space are transferred to corresponding grayscale images using the OTSU method, and the grayscale images are treated by nonlinear smoothing method to eliminate noise, and then the grayscale pictures are transferred to the two-color images using maximum variance between two groups to select the optimal sorting threshold value. In order to extract and identify the character of cotton seedling, the two-color images are treated refiningly, and removing spur treatment method is designed to eliminate the effect of the picture spur after refined on character identification. Further, the weight match board method is used, and match rule for preventing misjudge is designed. Using these series of method, the cotton seedling recognition is implemented finally, and it is explained that the method of using visual picture processing technology to recognize cotton seedling is feasible.
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  Detection of Cotton Leaf Curl Virus by Real-time Fluorescent PCR Combining with Magnetic Nanoparticles

  ZHANG Yong-Jiang, LI Ming-Fu, XIN Yan-Yan- , LI Gui-Fen, MA Jie, DENG Cong-Liang

  Cotton Science. 2013, 25(1): 90-94. https://doi.org/10.11963/cs130115

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  For developing a rapid and sensitive detection technique to detect cotton leaf curl virus (CLCuV) in order to prevent its introduction and spread, we have developed a Real-time TaqMan PCR combining with magnetic nanoparticles (MNPs) method. By using TaqMan probe combining with MNPs technique and coat protein gene as the target gene, the MNP fluorescent PCR detection method was established after specificity and sensitivity experiments. The sensitivity of the method was 525 fg·μL^-1 of DNA, which was same that of the conventional PCR gel electrophoresis method. The good specificity of this method was evaluated by applying the proposed method to detect five viruses, CLCuV, African cassava mosaic virus (ACMV), Tobacco streak virus (TSV), Cucumber mosaic virus (CMV) and Tomato spotted wilt virus (TSWV). The MNP fluorescent PCR method was rapid and accurate for CLCuV detection of field leaf samples and could reduce the risk of cross-contamination without any PCR post-processing, which is a feasible technique to routine testing assays.