15 January 2015, Volume 27 Issue 1

    

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  Development of EST-SSR Markers in Cotton Based on the Transcriptome Information of 10 Gland-related Materials

  Cheng Hailiang, Lu Cairui, Zou Changsong, Yu Daoqian, Jiang Pengfei, Yang Wencui, Zhang Youping, Wang Qiaolian, Song Guoli

  Cotton Science. 2015, 27(1): 1-8. https://doi.org/Y2015/V27/I1/1

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  Cotton is an important economic crop, however, the cottonseed is underutilized because of the presence of toxic gossypol, so low gossypol cotton breeding is an important part of the cotton breeding. The development of molecular markers and construction of high resolution genetic map will play an important role in research of genes relative to pigment glands, which are storage organ of gossypol, and other important traits. In this study, we developed EST-SSR markers based on the transcriptome information from 10 gland and glandless materials. 1546 EST-SSRs were obtained from 12895 unigenes more than 1 kb in length, which accounts for 11.99%, and the average distance of SSRs was 15.99 kb. Among the obtained EST-SSRs, dinucleotide and trinucleotide repeats were the dominant types, accounting for 30.85% and 48.97% respectively. AT/TA and GAA/CTT were the most abundant motifs respectively in dinucleotide and trinucleotide repeats. Total of 56 pairs of primers were designed and synthesized for the SSRs more than 20 bp and tested in the 10 materials to evaluate the application and polymorphism. 38 pairs of primers could produce clear bands in PAGE electrophoresis, accounting for 67.86%, and 9 EST-SSRs among them showed polymorphism, accounting for 23.68%. Further more, the functions and expression of the unigenes containing polymorphic EST-SSR were analyzed preliminarily. This study further proves that it is feasible to develop EST-SSR markers in cotton, and also basic work for the research of cotton high resolution genetic map and pigment gland related genes.
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  Generation of Transgenic Cotton(Gossypium hirsutum L.) with High-affinity K⁺ Transporter(AlHAK1) via Ovary-drip Method

  Liu Lingdi, Tang Hongliang, Dong Lijun, Zhang Shuling, Wu Lizhu, Du Huan, Gao Jiajia, Zhao Xiaoya, Liu Han, Jiang Xiao, Liu Jianfeng

  Cotton Science. 2015, 27(1): 9-15. https://doi.org/Y2015/V27/I1/9

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  The progress for development of transgenic engineering of cotton is restricted by plant regeneration difficulty, genotype-dependent response and a prolonged culture period and so on. In this study, a high-affinity K⁺ transporter(AlHAK1), isolated from a Aeluropus littoralis Parl., was introduced into cotton(Gossypium hirsutum L.) by a ovary-drip transformation method that is tissue culture-independent. The results showed that 44 out of 1006 transformed plants were kan-resistance and 35 were still positive by PCR detection, corresponding to a positive rate of 3.5%. Southern and Northern blot analysis confirmed that the AlHAK1 gene was correctly integrated into the genome and expressed in the transgenic plants. Under 0.05 mmol·L^－1 K⁺ concentrations stress, potassium contents in leaves and roots of T₁ generation transgenic plants were respectively 2-fold, 1.5-fold of that in the wild-type plants. While under 2.5 mmol·L^－1 K⁺ concentrations, there was no significant difference in potassium content between the transgenic plants and the wild-type plants. Under NaCl treatment, the germination rates of the transgenic cotton were remarkably higher than that of the wild-type plants within the NaCl concentration scope of 50～200 mmol·L^－1, especially under 150 mmol·L^－1 NaCl the germination rate of the transgenic lines was approximately 2.8-fold of that of wild-type plants The research findings may provide theoretical evidence for the extensive application of ovary-drip transformation system in production practice and important germplasm resources for breeding of new cotton variety adapting to K⁺ deficiency and salinity soil condittion.
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  Differential Response of High and Low Potassium Use Efficiency Cotton Genotypes to Drought and Low Soil Potassium Stress at the Flowering and Boll Setting Stage

  Wang Xiao, Chen Fang

  Cotton Science. 2015, 27(1): 16-21. https://doi.org/Y2015/V27/I1/16

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  To investigate the effects of soil water and potassium（K）on genotypic variation in photosynthesis, water and K use efficiencies, pot experiments were carried out with two cotton genotypes (a high K utilization efficiency cotton genotype and a low K utilization efficiency cotton genotype referred as HEG and LEG, respectively) and four treatments: optimum water and K (W1K1), drought stress(W2K1), low K stress(W1K2), drought and low K stresses(W2K2). The results indicated that soil drought had significantly negative effect on cotton net photosynthetic rate(P_n), stomatal conductance(G_s) and transpiration rate (T_r), then leading to falling of buds or bolls and decreasing the K use efficiency and yield. Application of K fertilizer improved cottons' P_n and water use efficiencies(WUE). There was a significant difference of WUE between genotypes. The WUE of LEG was notably and positively correlated with K applying, whereas HEG was on the contrary. Both P_n and T_r of HEG cotton decreased under low soil K condition, while WUE increased because the change of P_n was smaller than T_r. Higher WUE was conductive to higher absorption and transportation of K by increasing P_n and decreasing buds and boll falling ratio. Our findings will provide a new theoretical basis for further exploring of high K use efficiency mechanism.
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  Impact of Climate Change on Cotton-planting Zoning in Northern Xinjiang During 1961― 2012

  Li Jinglin, Pu Zongchao, Zhang Shanqing, Wang Mingquan, Li Xinjian, Xu Wenxiu

  Cotton Science. 2015, 27(1): 22-30. https://doi.org/Y2015/V27/I1/22

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  Based on the data of annual accumulated temperature of ≥10 ℃, July mean temperature and frost-free period from 52 meteorological stations in northern Xinjiang during 1961―2012, the spatial distributions of heat resources were analyzed by using mixed interpolation method based on ArcGIS, the change trends of heat elements were studied by using the linear regression method, and the variations in different periods of 1961―1970, 1971―1980, 1981―1990, 1991―2000 and 2001―2012 were analyzed also. Based on the above analyses, the cotton-planting climate zoning for northern Xinjiang in different periods were completed respectively with the climate indicators of cotton-planting zoning. And the variation of areas suitable, secondly suitable, risk and unsuitable for cotton in different periods were contrasted and analyzed. The main results were as follows: Heat resource was very obviously different in different regions of northern Xinjiang, generally, annual accumulated temperature of ≥10 ℃, July mean temperature and frost-free period were higher or longer in plain and basin areas than in mountain areas. In the background of global warming, the three indicators showed an increasing trend with rates of 75.657 ℃, 0.218 ℃and 4.36 d every 10 years respectively in recent 52 years. Under the joint effects of the above-mentioned climate factors, the areas suitable for cotton planting had a substantial expansion, and the areas secondly suitable and unsuitable for cotton reduced to different extents, but the areas risk for cotton planting did not change significantly. Compared with 1960s, the areas suitable for cotton planting increased 6.54164×10⁴ km², and the areas risk and unsuitable for cotton reduced by 0.99982×10⁴ km² and 5.28675×10⁴ km² respectively in 2001―2012. It is important to promote the development of cotton industry in the areas suitable for cotton planting increased in northern Xinjiang.
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  Cloning and Characterization of a Transcription Factor Gene GhMYB from Gossypium hirsuturm L.

  Yu Yuehua, Ni Zhiyong, Liang Xiaoli, Liu Zhenfang, Chen Quanjia, Gao Wenwei

  Cotton Science. 2015, 27(1): 31-38. https://doi.org/Y2015/V27/I1/31

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  MYB transcription factors, as one of the largest gene families in plants, play important roles in multiple biological processes. A gene coding for MYB, designated as GhMYB(GenBank No. HQ234875), was isolated from cotton. The full length GhMYB cDNA is 1264 bp, including a 250 bp 5'-UTR, an ORF（Open reading frame） of 774 bp, and a 240 bp 3'-UTR. This cDNA sequence encoded a polypepide of 256 amino acid residues with a predicted molecular mass of 28.867 kDa and a basic isoelectric point of 8.56, and two highly conserved SANT domains in the encoded putative protein. Furthermore, a length of 1355 bp sequence from genomic DNA of GhMYB was also cloned by PCR. The genomic DNA of GhMYB contains two exons and one intron. The deduced amino acid sequence had high homologies with MYBs from Theobroma cacao, Gossypium hirsutum and Prunus persica. Subcellular localization assay indicated the GhMYB:GFP fusion protein was targeted in the nucleus. SDS-PAGE indicated that the best expression quantity of the pET-28a-GhMYB was induced with 1.0 mmol·L-1 IPTG for 4 h at 37 ℃. Expression analysis with Unigene and GEO database showed that GhMYB gene was a boll specific expression gene and induced by waterlogging effect in root and leaf.
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  Dissipation Analysis of Flumetralin in Cotton Plants and Soil under Field Conditions and Risk Assessment for Dietary Residue Intake

  Wu Xujin, Wang Hong, Zhang Junfeng, Ma Huan, Zhou Juan, Wang Tieliang, Li Tong

  Cotton Science. 2015, 27(1): 39-45. https://doi.org/Y2015/V27/I1/39

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  In order to determine whether flumetralin usage was safe under the recommended dosage, we established an analytical method and investigated the dissipation dynamics and terminal residues of flumetralin in cotton crops and in soil under field conditions. The residue levels and dissipation rates of flumetralin in cotton and soil were determined by the gas chromatography using an electron capture detector (GC-ECD). At fortification levels of 0.05, 0.5 and 1 mg·kg^－1 in cotton leaves and in soil and of 0.1, 0.5 and 1 mg·kg^－1 in cotton seeds, recoveries were shown to range from 73.25 to 98.28 percent, with a relative standard deviation (RSD) of 1.68 to 9.93 percent. The limits of quantification (LOQ) were 0.05, 0.05 and 0.1 mg·kg^－1 for soil, cotton leaves and cotton seeds, respectively. The dissipation half-lives of flumetralin in cotton leaves and in soil were 0.81―3.7 days and 5.4― 8.7 days, respectively. The terminal residues at harvest in cotton seeds and in soil were lower than 0.54 mg·kg^－1 and 0.81 mg·kg^－1, respectively. The potential health risk induced by flumetralin was not significant based on supervised residue trial data. When the intervals to harvest were 20 and 30 days, there was no effect of dosage or frequency on the terminal residues; when the interval to harvest was 10 days, the dosage and frequency may have had an effect on the terminal residues. Our recommendations are that flumetralin (SC, 25%) be applied to wheat crops at a dosage of 450 g·hm^－2 using a pre-harvest interval (PHI) of at least 20 days between the last application and the harvest. These results would help the government to establish the maximum residue limit of flumetralin in cotton and to provide guidance concerning the proper and safe use of flumetralin.
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  Construction of SSR Fingerprinting Database of Standard Varieties on Cotton in DUS Testing

  Kuang Meng, Wang Yanqin, Zhou Dayun, Fang Dan, Ma Lei, Yang Weihua

  Cotton Science. 2015, 27(1): 46-52. https://doi.org/Y2015/V27/I1/46

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  Core SSR primers were used for constructing DNA fingerprint database of cotton DUS testing standard varieties based on fluorescence detection system and multiplex PCR (polymerase chain reaction) technology, and the genetic diversity was analyzed. Ten sets of 4 multiplex PCR combinations were established according to fundamental principle of multiplex PCR and characteristic of fluorescent system. DNA genetic analyzer was used for construction of fingerprinting database. Among the 30 varieties, 40 primer pairs had 146 allelic variations, and 3.65 allelic variations were detected by each SSR primer pair on an average with the range from 2 to7. The number of homozygous loci and heterozygous loci is almost the same. Clustering analysis indicated that sea island cotton (Hai7124) and upland cotton varieties were obviously divided into two categories. Xinluzao1 from Xinjiang cotton region differed from other upland cotton varieties. Multiplex fluorescence detection system has many advantages such as high-precision, high-throughput, high degree of automation compared to conventional silver staining system, especially for construction of large-scale fingerprint database. A primary idea about the application of molecular marker technology in cotton DUS testing by the construction of fingerprinting database of known varieties was proposed.
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  Allelopathic Effect of Aqueous Extracts of Eclipta prostrata L. on Cotton and Common Weeds in Cotton Field

  Yang Jinyan, Ma Xiaoyan, Jiang Weili, Ma Yajie, Ma Yan

  Cotton Science. 2015, 27(1): 53-61. https://doi.org/Y2015/V27/I1/53

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  This study was conducted to investigate the allelopathic effect of aqueous extracts of Eclipta prostrata L. on seed germination and seedling growth of cotton and six weed species in cotton field using Petri dishes. The results showed that aqueous extracts of Eclipta prostrata L. at concentrations from 0.01 to 0.08 g·mL^－1 strongly inhibited the germination of weed species, but only slightly inhibited those of CRRI 49 and CRRI 79. For different grass weeds, its inhibition effects to seed germination represented as Eleusine indica (L.) Gaertn. ＞Digitaria sanguinalis (L.) Scop. ＞Echinochloa crusgalli (L.) Beauv. For different broadleaf weeds, its inhibition effects to seed germination represented as Amaranthus retroflexus L. ＞Portulaca oleracea L. ＞Abutilon theophrasti Medic. The extracts stimulated growth of cotton and weed seedlings at lower concentrations, while inhibited their growth at higher concentrations, and inhibitory effects increased with the increasing concentration of the aqueous extract, with concentrations of 0.04―0.08 g·mL^－1 inhibiting radicle length and plumule length of A. retroflexus by 100%.
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  Effects of Continuous Cropping on Numbers of Fusarium oxysporum, Verticillium dahliae and Soil Bacterial Community in Cotton Field

  Liu Xiaolong, Ma Jianjiang, Guan Jizhao, Luo Jingjing, Luo Ming, Maidiniyeti·maimaiti

  Cotton Science. 2015, 27(1): 62-70. https://doi.org/Y2015/V27/I1/62

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   In this study, using selective cultivation medium to investigate numbers of Fusarium oxysporum and Verticillium dahliae, conducting PCR-DGGE (Polymerase chain reaction-denatured gradient gel electrophoresis) approach to analyze the diversity of soil bacterial community in the cotton field continuous cropping for different years. The results showed that continuous cropping cotton increased pathogen accumulation of Fusarium oxysporum and Verticillium dahliae in soil. The number of Fusarium oxysporum and Verticillium dahliae had similar change characteristics. With the extension of five to twenty continuous cropping years, the number of pathogen presented increasing trend, and the maximum was in continuous cropping 20 years; Over 20 years, the number of pathogen decreased. The quantity changes of Fusarium oxysporum and Verticillium dahliae had significantly positive relationship with Fusarium wilt and Verticillium wilt disease rate. DGGE patterns analysis indicated that continuous cropping could alter the composition, community structure and abundance of soil bacterial population in cotton field. The similarity of bacterial community structure between continuous cropping above 20 years and less than 20 years is only 32%, and the components of bacterial communities in the cotton field continuous cropping for 40 years were separated from others. For cotton fields continuous cropping from 5 years to 40 years, the Shannon-Wiener indices, evenness and richness indices of soil bacterial community structure diversity were decreased from 3.659 to 2.0854, 0.9504 to 0.7083, 47 to 14, respectively, and the trend dropped obviously with increasing of continuous cropping years.
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  Molecular Mechanisms of Premature Senescence in Cotton

  Kong Xiangqiang, Luo Zhen, Li Cundong, Dong Hezhong

  Cotton Science. 2015, 27(1): 71-79. https://doi.org/Y2015/V27/I1/71

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  Premature senescence is an abnormal phenomenon during plant growth and development in cotton, which can be due to its genetic background or external environment. Many senescence-associated genes(SAGs) are differently expressed during premature senescence. Most of the genes related to photosynthesis, anabolism of carbohydrates and other biomolecules are down-regulated, but those for catabolism of proteins, nucleic acids and lipids and the genes related to nutrient recycling, such as transport of amino acids, sugars, purines, pyrimidines and ions, are mostly up-regulated. Most of the hormone-related genes like ABA, ethylene, auxin, JA, and GA related genes are mostly up-regulated, while the cytokinin biosynthesis gene IPT is down-regulated during leaf senescence. Many NAC and WRKY transcription factor genes are up-regulated during premature senescence. Based on previous studies, the roles of the senescence-associated genes such as photosynthesis, biomacromolecule catabolism, nutrient recycling, hormone and transcription factor related genes have been reviewed to document the underlying mechanisms of premature senescence in cotton.
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  Mechanisms for Adapting to Waterlogging Stress in Cotton

  Zhang Yanjun, Dong Hezhong

  Cotton Science. 2015, 27(1): 80-88. https://doi.org/Y2015/V27/I1/80

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  Waterlogging is one of the major hazardous abiotic stresses which have dramatically negative impacts on cotton productivity. Waterlogging adversely affects plant growth and development of cotton, which may further result in considerable yield loss or crop failure when a moderate or a long and heavy waterlogging occurs. As one of the plant species with indeterminate growth habit, cotton has a series of mechanisms to protect itself from yield loss through adapting to waterlogging conditions. These waterlogging-adaptive mechanisms include escaping strategy, quiescent adaptive strategy and regeneration-compensation strategy. In this paper, we focused on the three strategies of cotton associated with tolerance and adaption to waterlogging. The adverse effects of waterlogging on plant growth and development, physiological traits, yield traits of cotton and measures in mitigating the adverse effects of waterlogging stress were also concisely reviewed.
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  Review of Researches and Utilizations on Germplasms with Different Leaf Shapes in Cotton

  Jiang Hui, Zhao Junsheng, Wang Jiabao, Chen Ying, Gao Mingwei, Wang Xiuli

  Cotton Science. 2015, 27(1): 89-94. https://doi.org/Y2015/V27/I1/89

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  We summarized the related study of genetics, breeding, cultivating and physiology about the shape of cotton leaf, and the problems were also analyzed in this paper. The acquired information could contribute to the further research of genetic theory of leaf shape and it's utilization in the breeding process.