15 March 2016, Volume 28 Issue 2
    

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  • Wang Yuanyuan, Liu Fang, Zhou Zhongli, Cai Xiaoyan, Wang Yuhong, Li Shaohui, Wang Chunying, Zhang Xiangdi, Wang Xingxing, Wang Kunbo
    Cotton Science. 2016, 28(2): 95-103. https://doi.org/10.11963/issn.1002-7807.201602001
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    While glyphosate makes weed management easy and efficient by controlling almost all emerged weeds, it severely influences the normal growth of cotton. The objective of this research was to evaluate natural glyphosate resistance by using tampons with different concentrations of glyphosate placed at specific positions on leaves of different plants and then rating the plants according to their reaction to glyphosate. The plants were graded into six levels of glyphosate resistance, with the range of the concentrations used (0.200% vs 0.006%) being 33.3 times higher than those that could be used in practice. In particular, Gossypium hirsutum races, whose resistance was distributed across all six levels, had more variability in glyphosate resistance than Gossypium arboreum. However, nearly 86% of G. hirsutum races had low or no tolerance to glyphosate while over 70% of G. arboreum plants were tolerant, revealing that G. arboreum is more glyphosate tolerant overall. This was also supported by evidence that only two accessions of G. hirsutum exhibited high resistance to glyphosate while five germplasms of G. arboreum were highly resistant. Furthermore, one semi-wild accession of G. hirsutum was susceptible to glyphosate, but no susceptible plants were found for G. arboreum. These materials provide a base for further in-depth study of the genetic mechanism of glyphosate-resistance, glyphosate-resistant cotton breeding, and internal resistant gene mining.
  • Wei Qichao, Zhang Rui, Sun Guoqing, Meng Zhigang, Zhou Tao, Zhao Leilin, Guo Sandui
    Cotton Science. 2016, 28(2): 104-114. https://doi.org/10.11963/issn.1002-7807.201602002
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    The aim of this study is to clone seed-specific promoter from Gossypium hirsutum. Using the genome sequence of Gossypium raimondii, primers were designed and synthesized. Upstream sequences of GhαGLOA, GhβGLOA, and GhβGLOB were amplified using total DNA of G. hirsutum as the template. Expression plasmids of GUS driven by the three upstream sequences were introduced into Arabidopsis thaliana. GUS activity was tested in seeds of transgenic Arabidopsis by fluorometric assay. The results showed that the three cloned sequences function as promoters, and that the promoter activity of GhαGLOA is significantly higher than that of GhβGLOA or GhβGLOB. Fluorometric analysis of GUS activity was carried out to examine the activity of these three promoters in different adult organs and tissues. Results showed that the expression level of GUS in roots, stems, leaves, and all other tissues examined, is very low, indicating that high levels of GUS activity were detected only in seeds. These results suggest that the transcriptional activity of promoters from GhαGLOA, GhβGLOA, and GhβGLOB was tightly controlled and highly seed specific.
  • Cotton Science. 2016, 28(2): 115-121. https://doi.org/10.11963/issn.1002-7807.201602003
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    To understand the genetic foundation of over-parent heterosis for boll weight in upland cotton(Gossypium hirsutum L.), we used mixed major-genes plus polygenes inheritance analysis of P1, F1, P2, and F2 populations to investigate inheritance of boll weight and boll shell ratio in three over-parent heterosis crosses L178 × L029 (Ⅰ), L178 × L057 (Ⅱ), and L029 × GP72 (Ⅲ) in 2013. The results showed that over-parent heterosis for boll weight and negative mid-parent heterosis for boll shell ratio were stable in the three crosses. Boll weight was significantly and negatively correlated with boll shell ratio in the F2 populations of crosses Ⅰ and Ⅱ. The two traits were controlled by two major genes plus polygenes, although the action modes for the major genes varied across crosses. The over-parent heterosis of boll weight was mainly controlled by the dominance effect of polygenes, although significant dominance-by-dominance interactions for major genes was observed in cross Ⅲ. The negative mid-parent heterosis of boll shell ratio was found to be associated with the dominance effect of major genes in crosses Ⅰ and Ⅲ, and with the dominance effect of polygenes in cross Ⅱ. Therefore, recurrent selection and modified backcrossing can be used to pyramid cotton novel alleles to improve boll weight and boll shell ratio.
  • Zhao Xiaojie, Mu Min, Lu Xuke, Yin Zujun, Wang Delong, Wang Junjuan, Fan Weili, Wang Shuai, Ye Wuwei
    Cotton Science. 2016, 28(2): 122-128. https://doi.org/10.11963/issn.1002-7807.201602004
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    To develop an expression system to study the function of the GhVP gene in cotton (Gossypium hirsutum), we successfully cloned GhVP and used it to construct a pBI121-GhVP∷GFP fluorescence expression vector. We transformed the vector into onion (Allium cepa) epidermal cells using gene gun technology and found that the GhVP protein was located on the cytomembrane. The pBI121-GhVP∷GFP vector was then transformed into cotton pollen. Detection of green fluorescence in the pollen indicated that the gene was successfully expressed from the pBI121-GhVP∷GFP vector. These results lay the foundation for the study of GhVP in cotton and the development of salt tolerance breeding.
  • Hou Yunpeng, Feng Hongjie, Cheng Wenhan, Zhang Xinyu, Sun Jie, Liu Yongchang
    Cotton Science. 2016, 28(2): 129-137. https://doi.org/10.11963/issn.1002-7807.201602005
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    To genetically transform cotton shoot tips, we established a biolistic genetic transformation system though the plant shoot tip culture technique and the gene gun-mediated transformation method, using the shoot tips of brown cotton Xincaimian No.5, a Gossypium hirsutum L. variety cultured in Xinjiang as materials. Several critical factors of the system were assessed, including the type of coagulant, the activated charcoal concentration, the kanamycin concentration, the penetration processing time, and the bombardment pressure and distance. Results showed that each factor clearly influenced the transformation rate of brown cotton shoot tips. An optimal biolistic genetic transformation technology system was determined: (1) shoot tips were cultured for 4 h before being bombarded in the osmotic medium; (2) the shoot tips were bombarded with helium pressure of 7.586 MPa (1100 psi), and a bombardment distance of 9 cm; (3) after culturing for 4 d in the recovery medium, shoot tips were moved to MSB screening medium containing 8 g·L-1 agar powder and 125 mg·L-1 kanamycin and then screened for 25 d to obtain the resistant germs; (4) then, 1 g·L-1 activated charcoal was added to the induction medium to induce rooting of the resistant germs to obtain resistant regenerating plants. After PCR and qPCR detections, 16 plants positive for the transgene were obtained from 5200 shoot tips, with a transformation rate of 0.31%. The establishment of this technology system lays a foundation for the genetic transformation of brown cotton and provides new materials for naturally-colored cotton breeding.
  • Xu Zhenzhen, Peng Zhen, Liu Jing, Shen Xinlian, Du Jianchang
    Cotton Science. 2016, 28(2): 138-143. https://doi.org/10.11963/issn.1002-7807.201602006
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    PIF-like elements are abundant in plant genomes and make up an important part of DNA transposons. In this study, we systematically studied the number, chromosome distribution, characteristic structure, and expression pattern of PIF-like elements in the Gossypium raimondii genome using genomic and bioinformatic methods. We also studied the types and functions of their adjacent genes. Four hundred and forty PIF-like elements that carried transposase and had clear insert positions were detected. These were distributed almost evenly on 13 chromosomes. One hundred and seventy-one adjacent protein-encoding genes with distances of less than 2 kb to the PIF-like elements were identified, 132 of which were annotated to the Gene Ontology database with a wide variety of functions. Some of the 440 PIF-like elements were expressed, with the expression patterns varying among different tissues. These results will provide a firm foundation for studying the evolution of genes and genomes mediated by PIF-like elements and the functions of protein-encoding genes in cotton.
  • Cotton Science. 2016, 28(2): 144-151. https://doi.org/10.11963/issn.1002-7807.201602007
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    To investigate cultivar differences in cotton seed vigor and the effects of different vigor determination methods on cotton seed vigor, we selected 17 cotton cultivars in which to measure seed vigor characteristics. We used simplified standard, low-temperature, and artificial aging germination test conditions, and analyzed the correlation between seed vigor and physical characteristics. Results showed many differences in seed vigor in the different cultivars, and that seed vigor characteristics were not consistent under the three germination test conditions. Seedling growth rate was mainly inhibited under low-temperature conditions, and seed germination percentage was mainly inhibited under artificial aging conditions. The correlation of the vigor index between simplified standard and stress germination was lower. High vigor cultivars CCRI 102, Xinluzhong 42, Xiangza 7, XG39K5, Tunza 6, and Wanza 9 were selected by cluster analysis of the three germination tests. In addition, the correlation between seed vigor and physical characteristics decreased significantly under stress conditions. Because healthy seed percentage and seed density were closely related to cotton seed vigor under different germination conditions, we also conclude that physical characteristics of the seeds can determine the level of seed vigor.
  • Wang Xiaoru, Dong Helin, Li Yongqi, Li Pengcheng, Liu Jingran, Liu Aizhong, Sun Miao, Zhao Xinhua, Li Yabing, Zhang Siping
    Cotton Science. 2016, 28(2): 152-159. https://doi.org/10.11963/issn.1002-7807.201602008
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    Hydroponics were used to explore the capacity of different cotton varieties to absorb potassium. Six cotton varieties were used to study the effect of different morphological and physiological characteristics on K uptake efficiency and K+ absorption kinetics parameters. The results showed that under low potassium stress, varieties such as CCRI 64, which had high K uptake efficiency, had the largest root dry weight, root volume, active uptake area, active specific surface area, and the strongest root vigor. The high K uptake efficiency variety CCRI 35 also had larger root dry weight, long and fine roots, the largest total root absorption area and total specific surface area, stronger root vigor, the highest K+ maximum absorption rate (Imax), and lower critical concentration(Cmin). CCRI 35 might, therefore, be tolerant of low K stress. Conversely, the K inefficient varieties Zhongzhimian No.2 and CCRI 50 had poor root morphology indicators, weaker root vigor, and smaller K+ maximum absorption rate. The research will not only help to clarify the reasons for resisting low potassium stress among cotton varieties but will also evaluate the adaptability of cotton varieties to low K.
  • Cotton Science. 2016, 28(2): 160-169. https://doi.org/10.11963/issn.1002-7807.201602009
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    This study explored the effect of the short-season and direct seeding cultivation on cotton yield levels at the Liuyang experimental base of Hunan Agricultural University in 2014. The "311" optimal regression design was performed to investigate the effects of sowing date, planting density, and nitrogen application rate on yield components and fiber quality of early-maturing variety JX0010. The effects of different treatments on the temporal and spatial distribution, cotton yield, and fiber quality of cotton were analyzed. The results showed that the temporal distribution of cotton boll number was highest in autumn, and the spatial distribution of cotton boll number was the highest in the middle and lower parts of the plant while the lateral distribution was mainly within-boll. According to the experimental results, the polynomial regression equation of three cultural factors; nitrogen application rate (x1), density (x2), sowing date (x3), with seed yield were established as follows: y=213.522+3.9331x1+6.414x2-21.4619x3-5.7541x12-4.321x22-7.5348x32-11.982x1x2-0.1645x1x3+3.5747x2x3. Single-factor effect analysis revealed that sowing date > planting density > nitrogen application rate. Results showed that the interaction effect between nitrogen application rate and planting density > planting density and sowing date > nitrogen application rate and sowing date. The optimum combination for high yield of seed cotton was a nitrogen application rate of 202.35 kg·hm-2, planting density of 6.2×104 plants·hm-2, and sowing date of May 19th. The highest cotton yield was 5510.25 kg·hm-2. The effect of cultivation factors on fiber quality indicated that micronaire > breaking elongation > breaking tenacity > fiber upper half mean length > uniformity index.
  • Cotton Science. 2016, 28(2): 170-178. https://doi.org/10.11963/issn.1002-7807.201602010
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    The endophytic bacterial communities in cotton roots infected with Verticillium dahliae were studied by extracting total DNA from the roots of healthy and infected plants. The V3 region of 16S rDNA was amplified and analyzed by denaturing gradient gel electrophoresis (DGGE). The 21 predominant bands were then sequenced. According to the phylogenic analysis, the samples at different periods were divided into two clusters. One cluster is the root samples from plants at the seedling and bud stages, the other is the root samples from plants at the flowering and boll setting stages. Overall, the diversity of endophytic bacterial communities decreased in the roots of cotton plants infected with V. dahliae. Sequence alignments in GenBank revealed that the majority of the bands (80.95%) belong to Proteobacteria and the rest (19.05%) were uncultured bacteria. The results showed that the diversity of the endophytic bacterial communities in cotton appeared to be influenced by V. dahliae.
  • Cotton Science. 2016, 28(2): 179-183. https://doi.org/10.11963/issn.1002-7807.201602011
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    While the draft genome of Gossypium raimondii provides a firm foundation for cotton fundamental research, it requires further work to make it complete and robust. Four-kilobyte-long sequences were selected from terminal sections of six of the longer chromosomes using the G. raimondii draft genome. After analysis of the selected sequences by bioinformatics, Chr2D was found to be a single-copy sequence without repetitive sequences. Obvious hybridization signals were detected in one terminal region of chromosomes 1 and 4 when sequence Chr2D was used as a probe and mitotic metaphase chromosomes of G. raimondii were used as target DNA. The size of the signal is significantly larger than a 4-kb sequence would account for while the signal brightness is fainter than that of normal hybridization signals. This indicates that Chr2D may not actually be a single-copy sequence and that the copy number of the sequence in one terminal region of chromosomes 1 and 4 may be huge. The brightness of the signal suggests that the distribution of repetitive sequence may be loose. The similarity of hybridization signals on chromosome 1 and 4 show that the pair of chromosomes may be homologous. These results will help complement the draft genome of G. raimondii.
  • Liu Suhua, Peng Yan, Peng Xiaofeng, Luo Zhen, Dong Hezhong
    Cotton Science. 2016, 28(2): 184-188. https://doi.org/10.11963/issn.1002-7807.201602012
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    This study examined the effects of regulated deficit irrigation and different plant densities on plant growth and development, yield, and fiber quality of cotton in dry land areas. A two-factorial experiment was conducted to determine the water-saving and yield-improving benefits of regulated deficit irrigation on the dry land of Xinjiang in 2014. Cotton was treated under different irrigation regimes (saturation irrigation, regular irrigation, deficit irrigation) and at different plant population densities (1.2×105, 1.8×105, and 2.4×105 plants·hm-2). Net photosynthetic rates were measured and the dry weight of vegetative and reproductive organs was determined and used to calculate the harvest index (seed cotton yield/final biological yield). Plant height, stem diameter, the number of bolls, lint percentage, and fiber quality were also examined. The results showed significant interaction effects of irrigation and plant density on biological yield, harvest index, and economic yield of cotton, but no effect on fiber quality. Regulated deficit irrigation significantly inhibited vegetative growth, but increased the harvest index. Appropriate increases in plant density significantly increased biological yield and the number of bolls per unit area under regulated deficit irrigation. Regulated deficit irrigation plus high plant density not only reduced water consumption by 20%, but also produced the same amount of cotton as normal irrigation at high or moderate plant density, or saturation irrigation at low plant density. Regulated deficit irrigation combined with increased plant density is, therefore, an effective alternative to save water without decreasing yield.