15 January 2016, Volume 28 Issue 1

    

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  Analysis of Phenotypic Variation in Fiber Yield and Quality in Segregative Populations of Hybrid Cotton CCRI 70

  Ye Zhenwei, Deng Xiaoying, Shi Yuzhen, Li Junwen, Liu Aiying, Gong Juwu, Gong Wankui, Shang Haihong, Chen Tingting, Ge Qun, Wang Yanling, Wu Yuxiang, Liu Zhi, Yuan Youlu

  Cotton Science. 2016, 28(1): 1-10. https://doi.org/10.11963/issn.1002-7807.201601001

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  The upland cotton cultivar CCRI 70, a transgenic hybrid with medium maturity and insect resistance, has a superior lint yield, fiber length, and strength characteristics. It was developed by crossing SGK 156, a cotton line with insect resistance, excellent yield, and comprehensive agronomic traits, with 901-001, an introgressive line with high fiber quality. Herein, we analyzed the lint yield and quality characteristics of the hybrid F₁ and its self-pollinated offspring F₂, F_2:3, F_2:4, and F_2:5. The F1 hybrid was shown with outstanding fiber quality, with average fiber length of 33.00 mm and fiber strength of 33.30 cN·tex^－1. The occurrence of fiber attributes exceeding those of the parents was common in the four segregative generations. The lint percentage was significantly negatively correlated with fiber length and strength, but positively correlated with micronaire value, indicating that it is difficult to improve fiber yield and quality simultaneously. Eleven excellent lines were selected from the F_2:5 progeny, of which four exceeded the control cultivar with respect to lint yield.Their average fiber length was ＞31.80 mm, the fiber strength was ＞31.0 cN·tex^－1, the boll weight was ＞6.26 g, and the highest lint percentage was 38.64 %. This shows that elite lines with a high fiber yield and quality can be screened from higher self-pollination generations of CCRI 70. Moreover,we obtained a large number of basic materials for the analysis of quantitative genetics, gene function, and gene pyramiding breeding programs in cotton.
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  Multiplex PCR for the Detection of Herbicide-Resistant Cotton, LLCotton25

  Yin Quan, Li Yi, Song Jun, Wang Dong, Zhang Fuli, Liu Wenjuan, Chang Lijuan, Lei Shaorong, Liu Yong

  Cotton Science. 2016, 28(1): 11-17. https://doi.org/10.11963/issn.1002-7807.201601002

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  This study aimed to establish a multiplex PCR detection method for the herbicide-resistant cotton line, LLCotton25. We selected the endogenous cotton reference gene SADI, the cauliflower mosaic virus promoter(P-CaMV 35S), Agrobacterium tumefaciens terminator(T-NOS), and the target gene(bar) as detection genes for multiplex PCR.We optimized PCR reaction conditions using sequences of specific Chinese national standard primers, and tested its specificity and sensitivity. The multiplex PCR detection system was then used to successfully detect the four genes SADI, P-CaMV 35S, T-NOS, and bar from the herbicide-resistant cotton LLCotton25, but not from other samples. These results indicate that this system is sufficiently specific to detect the herbicide-resistant cotton LLCotton25.
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  Cloning and Functional Analysis of an Alternative Splicing Sequence of Cotton EPSPS Gene

  Gong Yuanyong, Ni Wanchao, Guo Shuqiao, Shu Hongmei, Paerhatio Maimaiti, He Linchi

  Cotton Science. 2016, 28(1): 17-26. https://doi.org/10.11963/issn.1002-7807.201601003

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  Alternative splicing is a universal phenomenon in gene transcription to increase protein functional diversity, but it has not been widely reported in functional cotton genes. While cloning the cotton 5-enolpyruvylshikimate 3-phosphate synthase gene(EPSPS), we found an alternative splicing variant. Bioinformatic techniques analyses showed that the cDNA sequence of this alternative splicing sequence is 152 bp shorter than normal EPSPS cDNA, and would result in the introduction of an early stop codon. All introns in the normal EPSPS genomic sequence follow the commonsplicing rule of "GT-AG", while the last intron in the alternative splicing sequence follows the "AT-AC" splicing rule. Notable differences were identified between the predicted three-dimensional structure of the normal EPSPS cDNA sequence and the alternative splicing sequence. To determine the function of this alternative sequence, we constructed a prokaryotic expression vector in which the sequence was inserted into the pET32a vector, then transformed into host bacteria Escherichia coli BL21(DE3) ΔaroA.The alternative splicing sequence was shown to lack the function of the normal EPSPS cDNA sequence. This study enriches what is known about cotton EPSPS transcriptsand lays the foundation for further research into the transcriptional mechanisms of cotton EPSPS.
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  The Effect of High-Temperature Stress on Bt Protein Content and Nitrogen Metabolism in the Boll Shell of Bt Cotton

  Heng Li, Wang Jun, Abidallah Eltayib, Hua Mingming, Zhang Xiang, Chen Yuan, Chen Dehua

  Cotton Science. 2016, 28(1): 27-33. https://doi.org/10.11963/issn.1002-7807.201601004

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  This study explored the reason for observed low insect resistance under high temperatures in Bt cotton production.Two Bt cotton cultivars were selected as experimental materials, and their Bt protein content and the nitrogen metabolic physiology of the boll wall were investigated under the stress of high temperature(39 ℃) growth for 3, 5, and 7 days in an artificial climate chamber. The Bt protein content declined within 7 d of high temperature stress, with the greatest reduction observed within 3 d, especially at 1 d. Correspondingly, the boll wall soluble protein content, and activities of glutamate pyruvate transaminase and glutamate oxaloacetate transaminase reduced, while the free amino acid content, and protease and peptidase activities increased, with the biggest change also detected with in 1 d of the high temperature stress. These results suggest that insect resistance of the boll stage decreases markedly under a high temperature climate, indicating that suitable management strategies should be devised to prevent damage from boll worm and related pests.
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  Evolution Analysis of the Precision of Regional Cotton Variety Trials in China

  Xu Naiyin, Li Jian

  Cotton Science. 2016, 28(1): 34-41. https://doi.org/10.11963/issn.1002-7807.201601005

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  Evaluation of experimental precision is the foundation of scientific assessment to candidate varieties and test environments in regional crop trials. The distribution of one-year-one-site test precisions, the evolution dynamics of one-year-multi-location trials and the accuracy comparison among 12 major cotton characteristics were implemented to explore and evaluate the developmental status and evolution trends of the experiment precision in national cotton regional trials, and thus to provide a theoretical guidance for the reasonable layout and optimal design of national cotton regional trials’ scheme, by using the datasets collected from the national cotton regional trials in the Yangtze Valley, Yellow River Valley and the Northwest Inland cotton planting regions from 2000 to 2014. Research results showed that: (1) the percentage of one-year-one-site tests, those could discriminate the differences among cultivars up to 5%, 8%, 10%, 12%, 15% and 20%, were about 20%, 50%, 70%, 83%, 92% and 98%, respectively. (2) The precision evolution trend of one-year-multi-location trial increased year by year. The relative least significant difference at 0.05 level(RLSD_0.05) declined from 11.43% in 2000 to 7.1% in 2014 in nationwide, among of which RLSD_0.05 was continuously lower than 5% in recent successive 9 years in the Yangtze Valley, and 4 years in the Yellow River Valley, so that the trials in the two regions were accurate enough to reliably identify the 5% difference among cultivars. However, national cotton regional trials in the Northwest Inland region could reliably discriminate about 10% difference currently, although RLSD_0.05 value was also on the decline these years. (3) The experiment precision of 12 major cotton characteristics were significantly differenct, in which lint cotton yield and bolls per plant were lower in test precision, yield ratio before frost, lint percent, fiber length and growth period were of relativly high precision, while the rest of the traits were of medium accuracy.
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  Characterization and Expression Analysis of Five CBF/DREB Genes from Gossypium barbadense L.

  Li Yue, Dai Peihong, Liu Chao, Su Xiujuan, Kong Liying, Li Xiang, Li Caiyun, Liu Xiaodong

  Cotton Science. 2016, 28(1): 42-51. https://doi.org/10.11963/issn.1002-7807.201601006

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  Low temperature, drought, and salinity are major abiotic stresses that influence the survival, productivity, and geographical distribution of many important crops worldwide. The C-repeat/dehydration-responsive element binding transcription factors(CBF/DREB) are important proteins involved in the response to plant abiotic stresses. To better understand CBF genes, we identified five CBFs from island cotton(Gossypium barbadense L.) using abioinformatics approach. These genes (GbCBF1- GbCBF5) were shown to encode proteins that share a high similarity with plant cold stress-related CBF proteins, which contain the classic AP2 domain and two CBF signature sequences. Phylogenetic analysis revealed that the isolated cotton CBF genes could be classified into the A-1 subgroup of the DREB subfamily with CBF1-3 genes in Arabidopsis. Semi-quantitative reverse transcription PCR was used for the expression profiling of the five GbCBF genes under different abiotic stresses (200 mmol·L^－1 NaCl, drought, and 4 ℃). Two of the GbCBF genes were shown to be up-regulated in cotton plants under cold stress, while all the five genes were down-regulated under salinity and drought treatments. Our findings provide valuable information for further exploring the roles of the CBF genes in cotton development and in the response to stress.
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  Molecular Cloning and Expression Analysis of NAC Genes Specifically Expressed in the Fibers of Gossypium arboreum L.

  Wang Zhongna, Shang Haihong, Chen Tingting, Gong Wankui, Liu Aiying, Li Junwen, Shi Yuzhen, Gong Juwu, Ge Qun, Wang Lin, Li Wei, Li Pengtao, Bai Zhichuan, Yu Jie, Yuan Youlu

  Cotton Science. 2016, 28(1): 52-64. https://doi.org/10.11963/issn.1002-7807.201601007

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  A total of 29,729 fiber-related expressed sequence tags and the genome sequence of Gossypium arboreum L. were combined with NAC transcription factor family research findings from G. raimondii using bioinformatics to identify 143 NAC transcription factor genes from G. arboreum L. Quantitative real-time PCR was used to analyze the expression patterns of 30 of these genes, and the expressions of seven fiber advantage expressed genes were analyzed in G. hirsutum L., G. arboreum L., and G. raimondii. These seven genes showed different expression patterns in cotton plants with different genomes, suggesting that significant changes have taken place in these genes during the evolution of the cotton genome. The full-length cDNAs of three of these genes(designated GaNAC20, GaNAC09, and GaNAC69) were cloned successfully. Bioinformatic analysis showed that GaNAC20 and GaNAC09 are composed of three exons and two introns, while GaNAC69 has four exons and three introns. The encoded proteins have an NAM domain structure, which is typical of the NAC family of proteins. Multiple sequence alignment and phylogenetic tree analysis predicted the function of these proteins, and laid the foundation for the study of NAC transcription factors in the evolution of the cotton genome.
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  Development of EST-SSR and EST-InDel Markers Associated with Salt Tolerance in Upland Cotton

  Xu Peng, Cai Jihong, Yang Yang, Guo Qi, Zhang Xianggui, Xu Zhenzhen, Shen Xinlian

  Cotton Science. 2016, 28(1): 65-74. https://doi.org/10.11963/issn.1002-7807.201601008

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  With the development of cotton genomics and transcriptomics, cotton genetics and breeding would be enhanced by the identification of differentially expressed genes and the development of molecular makers based on candidate genes. The salt-tolerant cotton variety Miscott 7913-83 and salt-sensitive variety Su 12 were used as experimental materials in this study. RNA samples prepared from the roots and leaves of the two cultivars were pooled for transcriptome sequencing. Variations in gene expression were then examined after exposing the plants to 200 mmol·L^－1 NaCl for 12 and 72 h. A total of 3232 differentially expressed genes were then examined between Miscott 7913-83 and Su 12, and functional molecular markers such as EST-SSR and EST-InDel were designed by informatics tools according to differentially expressed gene sequences. Some SSR and InDel primers were randomly selected and further confirmed the accuracy. This research provides efficient methods for the rapid development of polymorphic markers in cotton. By focusing on functional molecular markers associated with salt tolerance, this should aid the improvement of salt tolerance by marker-assisted selection in upland cotton.
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  Phenotypic Analysis of Lint Yield and Fiber Quality Traits of Cotton Chromosome Segment Substitution Lines and F₁ Hybrids in Multiple Environments

  Li Botao, Shi Yuzhen, Gong Juwu, Li Junwen, Liu Aiying, Wang Tao, Shang Haihong, Gong Wankui, Chen Tingting, Ge Qun, Zhang Jinfeng, Wang Yongbo, Hu Yushu, Yuan Youlu

  Cotton Science. 2016, 28(1): 75-80. https://doi.org/10.11963/issn.1002-7807.201601009

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   A set of 82 cotton chromosome segment substitution lines(CSSLs) were selected and used to develop 41 F1 hybrids based on a random mating design. These F₁ plants were grown in five different environments to evaluate the lint yield and fiber quality traits of cotton. Positive control heterosis was detected for boll weight and lint yield in F₁ compared with CCRI 36, while mean fiber lengths and fiber strengths of F₁ and CSSLs were larger than those of CCRI 36 in all environments. Positive mid-parent heterosis(MPH) was detected for all yield traits of F₁ hybrids in all environments,while positive MPH was detected for fiber length, fiber strength, and fiber micronaire of F₁ in most environments.The MPH of F₁ hybrid fiber quality traits was smaller than that of yield traits. These results reveal the abundant genetic variations both in CSSLs and F1 hybrids, some of which are associated with a good lint yield and fiber quality. Our findings also show the feasibility of simultaneously improving fiber yield and quality using CSSLs, and the prospects of using heterosis and the genetic improvement of CSSLs in cotton breeding.
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  The Impact of Non-Bt Genetically Modified Cotton on the Community Diversity and Food-Web Structure of Arthropods

  Guo Jiahui, Ji Guozhen, Li Gang, Zhao Jianning, Yang Dianlin, Zhang Guilong, Yan Fengming, Xiu Weiming

  Cotton Science. 2016, 28(1): 81-86. https://doi.org/10.11963/issn.1002-7807.201601010

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  The influence of three genetically modified(GM) cotton lines, GM salt-tolerant cotton, GM disease-resistant cotton, and GM drought-resistant cotton, and their non-transgenic counterparts on the structure and diversity of arthropod communities and the food-web structure was investigated. No significant differences were found between GM cottons and their counterparts with respect to arthropod diversity, community evenness, or the dominant concentration index(P ＞0.05). There was also no significant difference in the basic characteristic values of the arthropod community foodweb between GM cottons and their counterparts(P ＞0.05). However, differences were observedbetween GM cottons and their counterparts regarding community similarity (73.18%-77.65%). These results show that the three GM cottons have no significant impact on the diversity or food-web structure of the arthropod community.
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  Responses of Cotton Phenology and Yield to Climate Warming in the Middle and Lower Reaches of the Yellow River Valley

  Zhang Lixia, Ma Gaigai, Ru Jingyi

  Cotton Science. 2016, 28(1): 87-94. https://doi.org/10.11963/issn.1002-7807.201601011

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  Exploring the effects of climate change on cotton phenology and yield could stimulate economic development and forecast cotton yield under future climate warming. In this study, a field warming experiment involving infrared radiators was conducted to investigate the responses of cotton phenology and yield to climate warming. A 0.6 ℃ rise of soil temperature was shown to advance the flower and boll stage by 3 days, and to extended the entire growth stage by 2 days. Warming also profoundly enhanced the leaf net photosynthetic rate at the flower and boll stage, and boll opening stage by 5.4% and 14.7%, respectively. During the growing stage, warming significantly increased the mean square and boll number of cotton by 7.4% and 7.2%, respectively. It also remarkablely enhanced the total plant biomass and distribution coefficient of the boll biomass by 32.2% and 11.0%,respectively, at the maturity stage, and significantly increased the boll mass per plant and above ground biomass per plant by 30.4% and 38.1%, respectively. Finally, warming also stimulated the cotton yield by 23.6%. This study suggests that climate warming may facilitate cotton yield in the region of the middle and lower reaches of the Yellow River.