15 March 2015, Volume 27 Issue 2
    

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  • The Oil and Fatty Acid Accumulation Patterns in Developing Cottonseeds of Xuzhou  142 and Its Fiberless and Fuzeeless Mutant
    Ma Jianjiang, Wu Man, Pei Wenfeng, Li Xingli, Huang Shuangling, Zhang Jinfa, Yu Jiwen, Yu Shuxun
    Cotton Science. 2015, 27(2): 95-103. https://doi.org/10.11963/issn.1002-7807.201502001
    Abstract ( ) Download PDF ( )   Knowledge map   Save
    The cottonseeds oil is one of the main edible oils and also one of the main raw materials in studies of biodiesel. However, there are few reports on the dynamic process of oil and fatty acids in developing cottonseeds. In this study, the accumulative process of oil and fatty acids were analyzed in developing cottonseeds(－3、－1、0、1、3、5、7、10、15、20、25、30、35 days post anthesis-DPA and maturation stage) from Xuzhou 142 and its fiberless and fuzzless mutant of Xuzhou 142. The results indicated that cottonseed oils of the two cotton lines from 20 DPA to the maturation stage were gradually increased; however, the cottonseeds in the mutant had significantly higher oil content than that of its wild type in each sampling date. Eight fatty acids were detectable in the whole development stages of the two genotypes' cottonseeds. Furthermore, C18:1 and C18:2 accounted for 72 percent of the total fatty acids in matural cottonseeds. After relevant analysis on the composition and content of fatty acids during the development process of cotton seeds, we found that C18:1 and C18:2 in Xuzhou 142 performed positive correlation to some extent, while that in mutant material presented significant positive correlation. Expression analyses indicated that the expression level of GhLPAAT1 and GhLPAAT2 had almost the same trend with the process of fat accumulation in these two genotypes' cottonseeds. qRT-PCR data revealed that GhLPAAT1 and GhLPAAT2 genes may play a role in oil accumulation of cottonseeds.
  • Association Analysis of Cottonseed Kernel Oil Content with SSR Markers in Gossypium hirsutum L. races
    Li Xiaona, Cai Xiaoyan, Kiflom Weldu Okubazghi, Wang Xingxing, Wang Chunying, Liu Fang, Zhou Zhongli, Li Cheng, Wang Yuhong, Zhu Shuijin, Peng Renhai, Wang Kunbo
    Cotton Science. 2015, 27(2): 104-110. https://doi.org/10.11963/issn.1002-7807.201502002
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    The study objectives were to estimate genetic diversity, population structure, and identify loci associated with cottonseed oil content in Gossypium hirsutum L. To do this, 89 G. hirsutum L. races accessions, and 215 pairs of SSR molecular markers were compared. Genetic diversity, population structure, and association mapping analysis were conducted using different software packages. The results showed that the polymorphism information content of the SSR markers ranged from 0.0222 to 0.7390 with a mean of 0.3452. The Shannon-Wiener diversity index(H') ranged from 0.0616 to 1.4412 with a mean of 0.5614. Population structure analysis divided the 89 accessions into two subpopulations. The 89 accessions had extensive genetic variation and a simple population structure, which revealed the natural population to be representative for further association analysis of quantitative traits in cotton. Association analysis showed that 12 markers were significantly associated with cottonseed oil content(P＜0.05) over 3 successive years. Of all the accessions, LAT141 had the highest average cottonseed Kernel oil content (41.90 %), revealing its potential to be exploited as a good source of high oil content. The findings of this study could be used in genetic improvement of cottonseed oil content of cultivated Gossypium species through marker assisted selective breeding programs, and could also be utilized as a reference database for mapping other agronomic qualities of semi-wild accessions.
  • Cloning and Expression Analysis of GhSKIP35 Gene in Gossypium hirsutum L．
    Liu kai, Zhang Huachong, Qi Fangjun, Zhang Wenwei, Jian Guiliang
    Cotton Science. 2015, 27(2): 111-117. https://doi.org/10.11963/issn.1002-7807.201502003
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    A new gene, GhSKIP35, was cloned from cotton cultivar Zhongzhimian KV1, which was selected by screening a suppression subtractive hybridization library of Zhongzhimian KV1. The open reading frame (ORF) was 1863 nucleotides (nts) long and encoded a protein of 620 amino acids. The putative protein contains a conserved ANK domain. The cloned gene was most closely related to Citrus sinensis SKIP35 protein by phylogenetic analysis. Real-time quantitative reverse transcription polymerase chain reaction showed that the GhSKIP35 gene was rapidly activated after inoculation with Verticillium dahliae. Gene expression reached a peak at 12 h post-inoculation (hpi), and then its expression gradually decreased within 24―48 h. It was assumed that GhSKIP35 might play an important role in the defense reaction against Verticillium wilt. Virus induced gene silencing was used to silence endogenous genes in resistant upland cotton cultivar Zhongzhimian KV1 by targeting a fragment of GhSKIP35. V991 was used to inoculate silenced cotton plants to identify disease resistance. The results showed that the disease indices of wild type Zhongzhimian KV1 and vector control plants were 9.38 and 11.54, respectively. The disease index of silenced GhSKIP35 plants was 50.75. These results confirmed that the GhSKIP35 gene was related to Verticillium resistance in cotton.
  • Association Analysis of Salt Tolerance with SSR Markers in Gossypium hirsutum L.
    Shao Bingxin, Wang Hongmei, Zhao Yunlei, Chen Wei, Guo Zhijun, Gong Haiyan, Sang Xiaohui, Ye Wuwei, Wang Junjuan
    Cotton Science. 2015, 27(2): 118-125. https://doi.org/10.11963/issn.1002-7807.201502004
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    In this study, 134 samples of upland cotton cultivars were selected. The seedling emergence rates were tested under NaCl stress at a concentration of 0.3%(mass fraction). SSR markers(74) were used to scan the genome of these materials. To find the SSR markers associated with salt tolerance, an association analysis between salt tolerance and SSR markers was performed using TASSEL 2.1, based on the analysis of genetic structure by Structure 2.3.4. The results showed the following. (1) The seedling emergence rates of 134 cultivars were significantly different, with 27 salt sensitive and 10 salt tolerant varieties identified. (2) One hundred forty-eight loci were found in 74 SSR markers involving 246 allelic variations, ranging from 2 to 7 with an average of 3.32 variations per SSR marker. The gene diversity ranged from 0.0295 to 0.4959 with a mean of 0.2897. The polymorphic information content ranged from 0.0290 to 0.3729 with a mean of 0.2381. (3) This natural population was classified into two subgroups by Structure 2.3.4. These two subgroups contained 89 and 45 samples, respectively. (4) Eight SSR sites associated with salt tolerance were found through association analysis, with the rate of explanation ranging from 2.91 to 7.82%, with a mean of 4.32%. The results could provide a reference for molecular marker assisted breeding of salt tolerance in cotton.
  • Genetic Analysis of Agronomic and Yield Traits of Upland Cotton under Continuous High Temperature and Drought
    Pei Xiaoyu, Zhou Xiaojian, Ma Xiongfeng, Tian Guiping, Wang Haifeng, Meng Qingqin, Zhang Wensheng, Zhou Kehai, Liu Yangai, Yang Daigang
    Cotton Science. 2015, 27(2): 126-134. https://doi.org/10.11963/issn.1002-7807.201502005
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    Heterosis is prevalent in nature. In this study, the NCII mating design approach, with nine upland cotton varieties (strains) forming 12 cross combinations, was used to analyze the main agronomic and yield traits in two ecological environments in 2013 with continuous high temperature and drought by the genetic model of additive-dominant with environmental interaction effects (ADE model). Genetic variance components were estimated, genetic correlations between all traits were analyzed and heterosis of F1 and F2 were predicted. The results indicated that agronomic and yield traits were affected by environmental conditions. Seed cotton yield, lint yield, boll number per plant and boll weight were controlled by dominant effects; lint percent was controlled by additive and dominant effects; and seed index was controlled by additive effects. Genetic correlation analysis showed that seed cotton yield and lint yield had positive dominant correlations with boll weight and lint percent; boll weight had a positive dominant correlation with lint percent, while lint percent had a negative dominant correlation and negative additive correlation with seed index. Heterosis analysis indicated that mean heterosis of boll weight, lint percentage, seed index, seed cotton yield and lint yield of CCRI 63 reached extremely significant levels, and the F3 of this variety had strong heterosis for lint yield.
  • Effects of Different Salt Control Measures after Saline Water Freezing Irrigation to Soil Water, Salt Dynamics, Cotton Emergence and Yield
    Feng Xiaohui, Zhang Xiumei, Guo Kai, Xie Zhixia, Liu Xiaojing
    Cotton Science. 2015, 27(2): 135-142. https://doi.org/10.11963/issn.1002-7807.201502006
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    Saline water freezing irrigation in winter is an effective way to plant cotton in the heavy saline land of northern China, where there is little fresh water to cultivate crops. After saline water ice melts in spring, mulching is necessary to maintain a low salt content in the surface soil. In this study we investigated the effects of different field soil salt control measures on the emergence rate of cotton seeds, growth characteristics and cotton yield. The results indicated that, in spring, film mulching and old film mulching could effectively reduce evaporation and salt accumulation at the soil surface. Straw mulching was the next most effective treatment, followed by liquid film, while tillage and intertillage were the least effective treatments. Emergence of cotton seeds was dependent on the salt content of the topsoil layer in May, which was closely related to soil moisture in March and April. Cotton yields were positively correlated with seed emergence rates. Cotton yields of fields mulched with plastic film and old film were the highest at 3226.13 and 3142.14 kg·hm－2, respectively, while cotton yields of fields mulched with straw (2029.81 kg·hm－2) and liquid film(782.79 kg·hm－2) were much lower. According to the cotton seed emergence rates and yields, this study found that old film mulching after the saline ice melted could effectively control salt accumulation and was also a time and labor saving measure.
  • Changes in Available Nutrients and Enzyme Activities in Soil of Transgenic(Bn-csRRM2) High-yield Cotton
    Zhao Yunli, Zhao Jianning, Li Gang, Xiu Weiming, Cao Xuan, Wang Hui, Ma Yan, Duo Li'an, Yang Dianlin
    Cotton Science. 2015, 27(2): 143-148. https://doi.org/10.11963/issn.1002-7807.201502007
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    This study monitored the changes in available nutrients and enzyme activities in the soils of transgenic(Bn-csRRM2) high-yield cotton and its parental isoline CRI 12 in different growth stages(seeding, squaring, flower and boll, and boll-opening stage) under field conditions. Results showed that ammonium nitrogen and nitrate nitrogen contents were not significantly different over time between cultivars, but the available phosphorus content was different over time between cultivars. The available phosphorus content was not significantly different between cultivars in squaring, flower and boll, and boll-opening stages. The ammonium nitrogen content was not significantly different between cultivars in seeding, squaring and flower and boll stages. The nitrate nitrogen content of transgenic high-yield cotton was significantly higher than that of conventional cotton in boll-opening stage and lower than conventional cotton in the other three growth stages. The urease and catalase activities were not significantly different between cultivars over time, but the alkaline phosphatase activity of transgenic high-yield cotton was slightly different from that of conventional cotton over time. Urease activity of transgenic high-yield cotton was significantly higher than that of conventional cotton in seeding, squaring, and boll-opening stages, but was not significantly different from that of conventional cotton in the flower and boll stage. There was no significant difference in catalase activity between cultivars in the seeding and boll-opening stages. Boll-opening stage was the only stage in which the alkaline phosphatase activity was not significantly different between cultivars. Our results indicated that the available nutrient contents and enzyme activities of soil were less affected by transgenic high-yield cotton, but mainly affected by the growth stage.
  • Activities of Detoxification Enzymes and Expressions of Related Genes in Spodoptera exigua Larvae Treated with Sublethal Concentrations of Emamectin Benzoate
    Dai Hanyang, Sun Yang, Bai Lixin, Zhao Jing, Xiao Liubin, Tan Yong'an
    Cotton Science. 2015, 27(2): 149-158. https://doi.org/10.11963/issn.1002-7807.201502008
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    To clarify the role of several important enzymatic detoxification steps in the formation of resistance against emamectin benzoate in Spodoptera exigua, the detoxifying enzyme activities and the expression of related genes were analyzed in S. exigua larvae treated with emamectin benzoate. When 3rd instar larvae of S. exigua were treated with emamectin benzoate at induction doses of LC5, LC20 and LC50 for 72 h, the mixed-function oxidase(MFO) activity and the expression of SeCYP450 genes were significantly increased(P＜0.01). The changes in activity or expression all increased at first and then decreased following the increasing doses of emamectin benzoate. The change in glutathione-S-transferase(GST) activity of S. exigua was similar to that for MFO, while the expression trends for SeGSTs and SeGSTs1 were different from the GST activity. Compared with the control, the expression of SeGSTs significantly increased in S. exigua treated with increasing induction doses of emamectin benzoate (P＜0.01), while expression of SeGSTs1 decreased. The esterase activities of S. exigua treated with three induction doses of emamectin benzoate all significantly decreased(P＜0.05), as did the expression of SeCarE(P＜0.05). Correlation analysis indicated that there was significant correlation between MFO, esterase activities and the expression of the corresponding genes SeCYP450 and SeCarE, respectively(P＜0.01). There was significant correlation between GST activities and SeGSTs expressions (P＜0.01), but no significant correlation between GST activities and SeGSTs1 expressions(P＞0.01). The MFO, GST and their related genes SeCYP450, SeGSTs and SeGSTs1 might be involves the evolution of resistance to emamectin benzoate in S. exigua larvae.
  • Changes in the Proteome of Susceptible Cotton Leaves after Inoculation of Verticillium dahliae
    Shen Fanrui, Zhang Wenwei, Zhang Huachong, Liu Kai, Jian Guiliang
    Cotton Science. 2015, 27(2): 159-165. https://doi.org/10.11963/issn.1002-7807.201502009
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    To explore the interaction mechanism between Gossypium hirsutum and Verticillium dahliae, total protein of Jimian 11 leaves inoculated with V. dahliae for 3 days was extracted to analyze proteomic changes. Twenty-two protein spots were found showing a change of more than 2-fold between the inoculated and the mock-inoculated samples, of which 10 were up-regulated and 12 were down-regulated. The up-regulated proteins included subunit beta of ATP synthase, RuBisCO activase 1, RuBisCO activase alpha 2 and fructose-1,6-bisphosphate aldolase. The down-regulated protein spots included chloroplast chlorophyll A-B binding protein, ribulose bisphosphate carboxylase big chain and ribulose bisphosphate carboxylase small chain. Real-time polymerase chain reaction of RBCS, cab and RCA1 genes showed that expression changes of the three genes were consistent at the mRNA and protein levels. From the analyses conducted, we hypothesized that the pathogen might interact with proteins participating in photosynthetic energy metabolism, disrupting cotton photosynthesis and causing wilting and withering of cotton leaves.
  • Diversity Analysis of Culturable Endophytic Bacteria with Antagonistic Activity against Fusarium oxysporum f. sp. vasinfectum or Verticillium dahliae from the Roots of Gossypium hirsutum
    Lin Ling, Wang Mingjiang, Zhou Yijun
    Cotton Science. 2015, 27(2): 166-175. https://doi.org/10.11963/issn.1002-7807.201502010
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    The aim of this study was to understand the diversity and antagonistic potential of cotton endophytic bacteria and to utilize endophytic bacteria for biocontrol of cotton vascular disease. Through antagonistic testing of endophytic bacteria against Fusarium oxysporum f.sp. vasinfectum and Verticillium dahliae, 87 strains of culturable endophytic antagonistic bacteria from the roots of Changkangmian and Simian 3 cotton cultivars were identified. Based on the amplified ribosomal DNA restriction analysis(ARDRA), these culturable endophytic antagonistic bacterial strains could be classified into 13 ARDRA groups. The 16S rRNA gene fragments of representative strains from different groups were sequenced and subjected to a BLAST search to identify similar strains. The results showed that the endophytic antagonistic bacteria most likely belonged to genera Bacillus, Brevibacillus, Burkholderia, Agrobacterium, Sinorhizobium, Acinetobacter and Pseudomonas. The endophytic antagonistic bacteria from the roots of Changkangmian belonged to genera Bacillus, Brevibacillus, Burkholderia and Agrobacterium, while the endophytic antagonistic bacteria from the roots of Simian 3 belonged to genera Bacillus, Burkholderia, Sinorhizobium, Acinetobacter and Pseudomonas. The most prominent genus from both varieties was Bacillus. BOX-PCR was used to study the diversity of the endophytic antagonistic bacteria and showed that a broad genetic diversity existed even within the same ARDRA group. Strains were tested for antagonistic activity against Fusarium oxysporum f. sp. vasinfectum and Verticillium dahliae and production of antibiotic substances including cellulase, chitinase, protease and siderophores. The results indicated that the biocontrol mecha nisms of these culturable endophytic antagonistic bacteria were different.
  • Cloning and Induced Expression Analysis of GhSAMDC2/3/4 in Cotton(Gossypium hirsutum L.)
    Wang Fanlong, Zhu Huaguo, Cheng Wenhan, Liu Yongchang, Cheng Xinqi, Sun Jie
    Cotton Science. 2015, 27(2): 176-183. https://doi.org/10.11963/issn.1002-7807.201502011
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    S-adenosine methionine decarboxylase(SAMDC) is one of three key enzymes in the polyamine synthesis pathway. GhSAMDC2/3/4(GhSAMDC2, GhSAMDC3 and GhSAMDC4) were acquired from Gossypium hirsutum L. by electronic cloning and reverse-transcription polymerase chain reaction(RT-PCR) technology. Sequence analysis showed that the gene cDNA contained two plant general SAMDC open reading frames(ORFs), the upstream ORF(uORF) and the main ORF(mORF). The mORFs of GhSAMDC2/3/4 were 1068, 1110 and 1032 bp, encoding 355, 369 and 343 amino acids, respectively. Cluster analysis showed that GhSAMDC2/3 and cacao tree SAMDC clustered within a class and GhSAMDC4 and AtSAMDC4 clustered within a separate category. Real-time PCR analysis showed GhSAMDC2/3/4 were expressed in all testing organization and fiber. GhSAMDC2 was obviously expressed in stem and late developed fiber. GhSAMDC2/3/4 were induced by abiotic stress, but GhSAMDC2 was strongly affected by low temperature and drought stress. GhSAMDC3 responded significantly to salt stress and GhSAMDC4 was strongly induced by ABA.
  • Effects of Sample Preparation on Quantitative Detection of Bt Protein in Genetically Modified Cotton
    Chang Lijuan, Liu Wenjuan, Zhang Fuli, Wang Dong, Yin Quan, Lei Shaorong, Guo Ling'an, Song Jun
    Cotton Science. 2015, 27(2): 184-188. https://doi.org/10.11963/issn.1002-7807.201502012
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    In order to evaluate the effects of sample preparation on quantitative detection of Bt protein in genetically modified cotton, several factors, such as different tissue sampled from leaves, different frozen storage time of the same samples and different particle size after grinding, were studied in this work using the transgenic cotton cultivars ZGK 9822 and GK l9 by ELISA. The results showed the content of Bt protein in vein was slightly higher than that in mesophyll and in petiole; Bt protein content per unit fresh mass weight and per unit total soluble protein decreased with the extension of frozen time. Significant differences were observed in Bt protein content per unit fresh mass weight between samples with frozen storage for 90 days and in samples with frozen storage for 60 days or 30 days, while no difference was found in Bt protein content per unit total soluble protein among samples with frozen storage for 30, 60 and 90 days; There was distinct impact on Bt protein content per unit fresh mass weight and per unit total soluble protein in samples with different particle sizes. Compared with samples with 50 mesh in size, Bt protein content per unit fresh mass weight in samples with 200 mesh in size increased by 56% and Bt protein content in unit total soluble protein increased by 38%.
ISSN 1002-7807　CN 41-1163/S
Superintendent: China Association for Science and Technology
Sponsored by: China Association of Agricultural Science Societies
Organized by: Institute of Cotton Research of CAAS
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ISSN 1002-7807　CN 41-1163/S
Superintendent: China Association for Science and Technology
Sponsored by: China Association of Agricultural Science Societies
Organized by: Institute of Cotton Research of CAAS
Copyright © Cotton Science, All Rights Reserved.
Tel: (0372)2525361/362/369　
Fax: (0372)2525361　
E-mail: journal@caas.cn
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