15 September 2012, Volume 24 Issue 5
    

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  • PENG Ren-Hai, LIU Yu-Ling, LIU  Fang, WANG Chun-Ying, LI Shao-Hui, ZHANG Xiang-Di, WANG Yu-Hong, WANG Kun-Bo
    Cotton Science. 2012, 24(5): 379-385. https://doi.org/10.11963/cs120501
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    In order to obtained the high-quality well-differentiated mitotic metaphase chromosomes with clear background for laser microdissection in cotton, integrated mothed of pre-hypotonicity, enzymolysis, post-hypotonicity and squashed on mitotic metaphase root tip cell was adopted, the high quality preparations of Gossypium arboreum were obtained, and the single chromsome laser cutting technique in cotton was constructed. Amplified single chromosome production was verified by the integrated method of simple sequence repeats(SSR) primer amplification, Southern blotting and fluorescence in situ hybridization (FISH). The 7th chromosome library of Shixiya No. 1, a variety of G. arboreum was constructed. The library contains 1.38×105 clones, which almost cover 1 time of the chromosome length. The inserts averaged 550 bp, varied from 150 bp to 1000 bp. And the titer was 1.3×106 pfu·mL-1, rate of no-load was 1%, and the ration of single and low copy sequences were over 59%. The library will facilitate the specific probe screening, gene cloning and sitting, genetic map saturation on this chromosome.
  • WEI Jun, WANG Wang-Hua, FAN Hong-Hong, LI Ting-Chun, CAI Yong-Ping, LIN Yi
    Cotton Science. 2012, 24(5): 386-392. https://doi.org/10.11963/cs120502
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    The objective of this study was to determine temporal expression profiles of transcripts during cold acclimation of cotton plants using cDNA-AFLP. The upland cotton line, ZongCaiXuan No.1, was used for expression analysis. After being screened with 64 primer combinations, 2321 bands were amplified and 34 polymorphic transcript-derived fragments(TDFs) were identified. According to differences in gene expression patterns, 22 polymorphic TDFs were isolated and cloned. Among the 22 TDFs, 18 were found to be related with the low temperature treatment, including two with up-regulated expression, nine with down-regulated expression and seven that were transiently expressed. In addition, there were four TDFs expressed specifically after a two-day recovery under normal conditions. An NCBI Blast search showed that 21 TDFs were homologous to functional genes, including a hypothetical protein, a transposon, a peroxisome proliferator activated receptor and stress defense genes, but the function of other one TDF remains unknown.
  • LI Qiong, QU Yan-Ying, YANG Ting, WANG Li-Ping, LIU Zheng, ZHOU Li-Rong, CHEN Quan-Jia
    Cotton Science. 2012, 24(5): 393-398. https://doi.org/10.11963/cs120503
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    By using two different methods, the floral spray and the plasmid injection, the Bt gene was introduced into cotton Junhai 1(Xinjiang sea island cotton) . T1 and T2 transformed progenies were confirmed by field screening, PCR amplification, Southern bolt hybridization and conformity analysis of Mendelian inheritance. The results showed: The boll rate of T0-generation transformed by the floral spray is 8.3 percent approximately higher than that of the plasmid injection. In this study, it has been proved that transgenic plants can be obtained through each of the two methods, and the transformation efficiency of the former is 3.4% higher than that of the latter; the floral spray is more in line with Mendelian inheritance, and probability of single copy is higher. So, the better one is the floral spray.
  • DI Li-Chao, ZHANG Yong-Jiang, LIU Lian-Tao, SUN Hong-Chun, ZHU Xiu-Jin, LI Cun-Dong
    Cotton Science. 2012, 24(5): 399-405. https://doi.org/10.11963/cs120504
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    Bt-transgenic cotton plants with two different sowing dates were treated normally (CK), or by removing one(R1), two (R2) or three basal fruiting branches(R3) to study the effects of early fruiting branch removal on growth, development and senescence processes in a two-year experiment. The results showed that the removal of early fruiting branches delayed the process of growth and development(there was no difference between R1 and CK in the total number of fruiting branches, but R2 and R3 were significantly lower than CK), but had no obvious effects on the total fruiting nodes. Early fruiting branch removal significantly increased the LAI and dry matter accumulation in plants of the first sowing date over two years(R1, R2 and R3 increased the dry matter weight on average by 13.5%, 17.5% and 19.0%, respectively, compared with CK), but there was no significant difference in plants of the second sowing date. For plants of the first sowing date, early fruiting branch removal significantly decreased the abscission and rotten boll rates and promoted seed yield over the two year period. R1, R2 and R3 decreased the abscission rate by 5.9%, 6.4% and 9.5%, respectively, and rotten boll rate by 3.8%, 3.1% and 4.3%, and the R1 and R2 treatments enhanced the seed cotton yield by 9.6% and 8.1%, respectively. There was no significant yield increase in plants of the second sowing date; there was even a decrease for some treatments.
  • XIN Cheng-Song, LUO Zhen, WU Zhen-Mei
    Cotton Science. 2012, 24(5): 406-413. https://doi.org/10.11963/cs120505
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    The physiological mechanisms of differences in salt tolerance among four genotypes of insect-resistant cotton were investigated to guide the appraisal of salt-tolerant cultivars and cotton production in saline fields. Sand and soil culture experiments with different salinity levels were conducted with strongly salt-tolerant cultivars(LK536 and the promising selection SCRC18) and weakly salt-tolerant ones(LK1258 and the promising selection 33B) of Bt transgenic cotton. Changes in some physiological and biochemical indices at the seedling stage were compared. The results showed that the comprehensive indices for salt tolerance of LK536 and SCRC18 were significantly higher than those of 33B and LK1258. In seedling leaves of strongly salt-tolerant cultivars, K+ content was significantly higher, while Na+ content was significantly lower; malondialdehyde content was also significantly lower, but the contents of free proline, soluble sugars, and carotenoids were all significantly higher than that in the seedling leaves of weakly salt-tolerant cultivars, respectively, under NaCl stress. The peroxidase activity, contents of chlorophyll a, b and a+b, and net photosynthetic rate of strongly salt-tolerant cultivars were substantially higher than those of weakly salt-tolerant cultivars at the seedling stage under NaCl stress. Further analysis indicated that the salt tolerance of cotton varied among genotypes. The variation in salt tolerance among genotypes was related to differences in the contents of malondialdehyde, free proline, soluble sugars, and carotenoids; in peroxidase activity;  and in the accumulation of salt ions in leaves.
  • LI Jie, FAN Shu-Li, SONG Mei-Zhen, PANG Chao-You, YU Shu-Xun
    Cotton Science. 2012, 24(5): 414-419. https://doi.org/10.11963/cs120506
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    GhSPL3, an SPL transcription factor, was cloned from upland cotton by bioinformation and RT-PCR technology. The sequence accession number is JN795132 in GenBank. The ORF length of GhSPL3 is 426 bp which encodes 141 amino acid residues. Bioinformatics analysis showed that GhSPL3 contains a typical SBP structure domain and a nuclear localization signal; Evolutionary tree analysis revealed that GhSPL3 and AtSPL3 were clustered in same group, thus GhSPL3 and AtSPL3 might have similar structure and function. Subcellular localization results showed that the signal of GhSPL3 was localized in the nucleus. Quantitative RT-PCR results showed that GhSPL3 expressed in all tissues, but the expression levels were different. The expression of GhSPL3 in the flowers was the highest, and the lower was that in shoot apexes and in stem, followed by that in the root and in the leaves. Among different development stages of shoot apex, the expression of GhSPL3 was the highest at the third true leaf expansion stage in shoot apex. The result of Quantitative RT-PCR indicated that GhSPL3 might play an important role in bud differentiation, the transition of the growth phase and flower formation.
  • TIAN Shao-Ren, NIE Tai-Li, WANG Meng-Liang, YANG Jun
    Cotton Science. 2012, 24(5): 420-426. https://doi.org/10.11963/cs120507
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    The mechanisms of antagonists against fungal pathogens have become increasingly important in biological control in recent decades. In this paper, the mechanisms of antagonism, competition, and systemic-resistance induction in the bacterium strain C-02 against Verticillium wilt of cotton were studied by double culture, liquid fermentation, and inducing cotton seedlings, respectively. The results indicated that an antifungal protein from strain C-02 could destroy the cell wall of pathogen strain VD-11 and inhibit its toxin secretion. The results also showed that strain C-02 could colonize the rhizospheric soil of cotton fields better than VD-11, and they had the advantage in carbon resource use. When cotton leaves were treated with a bacterial culture of C-02, PAL, POD and PPO activity was quickly enhanced and remained at high levels for a long time. In conclusion, the antagonistic mechanisms of C-02 against Verticillium wilt of cotton form a comprehensive behavior including antagonism, bacteriolysis, competition, and systemic-resistance induction.
  • WANG Zi-Sheng, JIN Lu-Lu, ZHAO Wen-Qing, WU Xiao-Dong, ZHAO Zhi-Qiang, WANG You-Hua, ZHOU Zhi-Guo
    Cotton Science. 2012, 24(5): 427-434. https://doi.org/10.11963/cs120508
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    To determine the critical nitrogen(N) concentration dilution curve for cotton grown in the extremely early-maturation region of northeast China, field experiments were carried out in Liaoyang (41°26'N, 123°14'E) with cultivars of Liaomian 19 and NuCOTN 33B. Three plant population densities (7.5×104, 9.75×104, and 12×104 plants·hm-2) and five nitrogen rates (0, 120, 240, 360, and 480 kg·hm-2) were applied. The results showed that the relationship between shoot dry matter and critical N concentration could be described by a power equation(N=aW-b). The coefficients a and b of the critical N dilution curve for cotton depended on cotton genotype and plant density. At higher plant densities, cotton plants had a higher capacity for N accumulation in shoot biomass than at lower densities. Variety NuCOTN 33B had a higher capacity for N accumulation in shoot biomass than did Liaomian 19 at the same plant density. Based on the critical N dilution model and the allometric relationship between N uptake and dry matter accumulation in shoots, models for potential growth and yield were developed for N nutrition index(NNI) and for N demand at different growth stages. According to the allometric growth coefficient, NNI, and N accumulation rate below the critical N concentration, we confirmed that the optimal rate of N application at a density of 9.75×104 plants·hm-2 was 240 kg·hm-2, which was lower than that at densities of 7.5×104 or 12 ×104 plants·hm-2 in the cotton extremely early-maturation region in northeast China.
  • LIU Zhang-Wei, FENG  Juan, FAN Xin-Qi, XU  Peng, ZHANG Xiang-Gui, SHEN Xin-Lian
    Cotton Science. 2012, 24(5): 435-443. https://doi.org/10.11963/cs120509
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    In this study, we used a salt-tolerant diploid G. aridum species and the salt-sensitive G. hirsutum cultivar Sumian 12 to investigate differential expression in the presence of salt stress. Using cDNA-AFLP, 25 transcript-derived fragments(TDFs) were isolated and confirmed to be present only in salt tolerant species G. aridum under salt-stressed conditions. BLAST analysis with sequences assembled using an in-silico approach demonstrated that 23 of the cDNA fragments had homology to known proteins. The up-regulated genes were mainly involved in ion transport, ROS scavenging, cell signaling, cell division, transcription regulation, membrane protection, and penetration regulation. Quantitative real-time PCR(qRT-PCR) was used to analyze the expression patterns of nine of the 23 TDFs at different stages of salt stress in the tolerant species G. aridum. Their expression showed a significant increase and reached a peak between 12–24 h of stress. These TDFs may serve as useful resources for future research on molecular mechanisms of salt stress response in cotton.
  • YANG Wei-Hua, XU Hong-Xia, WANG Yan-Qin, ZHOU Da-Yun, KUANG Meng, FENG Xin-Ai
    Cotton Science. 2012, 24(5): 444-450. https://doi.org/10.11963/cs120510
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    More than 600 cotton varieties(hybrids) were examined and approved by the National and Provincial Committees of Crop Variety Identification from 2006-2010. For the white-fiber varieties of upland cotton, the upper-half mean length ranged mainly between 29-30 mm; the uniformity index was mainly in the range of 84%-85%; fiber strength varied mainly from 28-30 cN·tex-1; and micronaire values were mostly over 4.3. Over 90% varieties met the standards for high-quality cotton. Most of the white upland cotton varieties were better than the varieties released in the United States with respect to upper-half mean length, uniformity index, and fiber strength, but the micronaire values were too high. This paper discussed some problems and potential suggestions for improvement.
  • XIONG Zong-Wei, WANG Xue-Jiao, GU Sheng-Hao, MAO Li-Li, ZHANG Li-Zhen, ZHOU Zhi-Guo-
    Cotton Science. 2012, 24(5): 451-460. https://doi.org/10.11963/cs120511
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    This paper reviewed cotton fiber quality and its evaluation method in china. We discussed research progress of cotton color feature index e.g. fiber light reflection ratio (Rd) and fiber yellowness (+b), and compared with traditional fiber classification such as visual grade, fiber length, strength and fineness. We also analyzed the effects of ecological climatic factors on the cotton fiber quality, and the correlation between cotton fiber quality index and cotton color feature index. The results indicated the importance of application of cotton color feature in demand of standard cotton quality test. We proposed to build a new classification system of cotton quality, using GIS platform, ecological index and cotton color feature to evaluate and forecast the spacial distribution of cotton fiber quality.
  • ZHOU Fang-Fang, LI Zhi-Fang, FENG Zi-Li, SHI Yong-Qiang, ZHAO Li-Hong, LI Cai-Hong, ZHU He-Qin
    Cotton Science. 2012, 24(5): 461-467. https://doi.org/10.11963/cs120512
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    Agrobacterium tumefaciens-mediated transformation(ATMT) has become an important tool for strain improvement, new genes' tagging and target gene's screening, isolating and cloning. The advantages of this method are simple operation, a wide host range, highly efficient transformation, and a high frequency of single inserted T-DNA. Mutant libraries of many fungi with high capacity and quality have been constructed by ATMT. The screening and identification of related functional genes will be the following research focus on the basis of these mutant libraries. This research will provide a basis for further studies on pathogenic mechanisms and will aid disease-resistance breeding.
  • GUO Zhong-Jun, CHEN Quan-Jia, XU Chun-Hua, WANG Xi-Dong, SU Xiu-Juan, QU Yan-Ying
    Cotton Science. 2012, 24(5): 468-472. https://doi.org/10.11963/cs120513
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    Using cotton(Gossypium hirsutum L.) leaves as experimental materials, the present study aimed at solving the problem of disturbance by secondary metabolites in the materials,such as chromatophores, phenols and quinones, and optimizing the protein extraction method, lysis buffer reagent, protein loading amounts and other factors. The results showed that optimal cotton leaf protein extraction was obtained by the improved TCA/acetone method, with lysis buffer containing 7 mol·L-1 Urea, 2 mol·L-1 Thiourea, 4% CHAPS, 80 mmol·L-1 DTT, 1 mmol·L-1 PMSF, and 0.3% Bio-Lyte, a protein loading amount of 600 μg, and an IPG (pH 4-7, 24 cm). Under these conditions a good protein pattern with protein spots distributed equally, of high quality and repeatability, was obtained. This system will establish a foundation for cotton proteomic research.