15 July 2009, Volume 21 Issue 4
    

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  • TONG Xu-Hong, WU Yu-Xiang, ZHU Shui-Jin
    Cotton Science. 2009, 21(4): 259-264.
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    5-Enolpyruvylshikimate-3-phosphate (EPSP) synthase is an enzyme of the shikimate pathway. It is the target enzyme of nonselective herbicide glyphosate. This enzyme is located in the chloroplasts of the higher plants. Primers were designed according to the assembled ESTs of Gossypium hirsutum, and a 1834 bp cDNA fragment of G.hirsutum cv. Coker 312 was obtained in this experiment. The frafment contains an ORF comprising of 1565 nucleotides which encodes 521 amino acid. A comparison of the deduced amino acid sequences of cotton and other higher plants genes reveals highly homology over the length of the entire mature protein. The site of transit peptides cleavage is deduced from a comparison to the known cleavage site in other plants. The enzyme, including 74 amino acid chloroplast transit peptides and 447 amino acid mature proteins, comprises conserved region of PEP binding sites and the characteristic sequence of catalytic activity sites. Semi-quantitative analysis of EPSP synthase expression showed that the gene expressed extensively in cotton roots,stems,leaves, and other organs, and leaf is the most highly expressed tissue. A genomic DNA sequence with 3344 bp clone containing an EPSP synthase was isolated from the upland cotton in this experiment, and the EPSPS gene contains eight introns and seven exons.
  • Article
  • DOU Li-Ping, WANG Qing-Ya, TANG Can-Ming
    Cotton Science. 2009, 21(4): 265-270.
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    The double recessiveness nuclear male-sterile line ms5ms6 of upland cotton (Gossypium hirsutum L.) has been used in cotton seed production, but there is no report on its abortion stage and character. The result of embryology study showed the abortion stages were the development periods of pollen mother cells and microspore, which was the main abortion stage observed using paraffin slice technique under light microscope. The abortion characters of the double recessiveness nuclear malesterile line ms5ms6 of upland cotton were: low frequency of nucleolus penetration through nucleus envelop and cell wall and part of pollen mother cells appeared in the shape of lunule; microspore adhesion and few of pentads were found during the periods of microspore; there was no spinescent protuberance on the pollen wall, finally the microspore appeared abnormal and breakdown; there was no difference between fertile anther and sterile anther about the tapetum but the function of tapetum producing corpus callosum is abnormal.
  • ZHAO Chang-Yan, WANG Sheng-Fen, HAN Gai-Ying, CHI Ji-Na, ZHANG Gui-Yin, MA Zhi-Ying
    Cotton Science. 2009, 21(4): 271-274.
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    The influences of Agrobacterium concentration,infection time,the time and the temperature of co-cultute on the induction ratios of kanamycin-resistant calli were studied by Agrobacterium-mediated transformation using the hypocotyls of Jiwu 2031 as explants.The factors were optimized using orthogonal design.The results showed that the transformation ratio was the highest under the regime of OD=0.3 of bacteria concentration,15 min of infection time,24 ℃ of co-culture temperature and 24 h of co-culture time.High frequency of kanamycin-resistant calli formation was observed when infecting hypocotyls of Coker 312,Zhong 521 and Nongda 94-7 using the optimized combination of factors,and the induction ratios reached 93.85%,86.72% and 85.83%,respectively.
  • WANG Yi-Xue, SUN Yi, TIAN Ying-Chuan, WU Jia-He
    Cotton Science. 2009, 21(4): 275-278.
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    The total RNA of Gossypium hirsutum was extracted by modified CTAB method and the open reading frame of enolase gene from Gossypium hirsutum was cloned by RT-PCR,named GhEno, which was 1338 bp long and encoded 445 amino acids.The GhEno gene was registered in GenBank with the access number of  EU169604.Sequence analysis alignment indicated that GhEno gene had high homology with those of other plants. The phylogenetic tree analysis showed that the GhEno was clustered in the same group with Hevea brasiliensis and Ricinus communis. Semi-quantity RT-PCR analysis indicated that expression of GhEno was detected in roots, stems and leaves,however it was stronger in roots than in leaves and stems.
  • DONG Zhang-Hui, SHI Yu-Zhen, ZHANG Jian-Hong, WANG Shu-Fang, LI Jun-Wen, LIU Ai-Ying, TANG Shu-Rong, CHU Ping, YUAN You-Lu
    Cotton Science. 2009, 21(4): 279-283.
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    TG41 and sGK156, two commercial cotton cultivars (lines), and HS427-10, 7235 and 0-153, three elite fiber quality germplasm lines, were used as parents to develop three double-cross combinations, (sGK156×HS427-10)×(0-153×7235), (TG41×HS427-10)×(0-153×7235) and (sGK156×0-153)×(sGK156×HS427-10). Three SSR markers, NAU1043, MUSS497 and CM67, which were linked with QTLs of fiber length and derived from 7235 or 0-153, were used to study the effect of molecular marker-assisted selection and pyramiding breeding in the three combinations, respectively. As a result, the fiber length showed significant differences between plants with and without marker in the three combinations. This indicated that the effects of the three QTLs for fiber length were stable in different genetic backgrounds and different generations. When selected the plants with two or three markers, the fiber length of plants with two or three markers were better than that of the plants with only one marker, and the fiber length was the best in the plants with all the three markers. In conclusion, it is effective to increase fiber length through molecular marker-assisted selection and the effect may be increased when more QTLs are  pyramided in plants. It is necessary to develop stable recombinant inbred lines of advanced generations which pyramid several fiber-related genes to study the effect for pyramiding more genes.
  • LIN Hai-Rong, LI Zhang-Cheng, ZHOU Qing-Bo, Lü Xin
    Cotton Science. 2009, 21(4): 284-289.
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    Frostbite occurrence lies mainly on condition of weather and vegetation.Frostbite can be monitored by remote sensing according to the difference of vegetation index,which has higher spatial and temporal resolution, and to the canopy temperature,which has higher precision of temperature retrieval.In addition,considering the aerosol and humidity effect on the temperature retrieval and the quality of the remote sensing image,monitoring the frostbite by remote sensing is rather complicated and difficult.North Xinjiang is the main production zone of cotton in China,where cotton is influenced seriously by later spring frost or early autumn low-temperature injury.Remote sensing is proved feasible in monitoring crop growth,especially after stresses.But monitoring frostbite of cotton with remote sensing has been rarely studied so far.So combining with the statistical data of agriculture disaster and crop development,according to the difference of NDVI and canopy temperature (CT), the remote sensing was applied to three temporal Landsat ETM images in 2000-08-07,2001-06-07 and 2001-08-10,the remote sensing monitor the frostbite of cotton happened in Xinjiang Shawan and Shihezi county.The results showed the pixels of the absolute difference of NDVI fallen was 67.8%,the year fallen great than 0.2 was 17%,the percent of difference NDVI decreased within 20%,apparently,the crop status of cotton in 2001 was worse than the year 2000's.The scatter plot between canopy temperature and NDVI showd the remarkably negative correlation,the correlation coefficient was -0.63.In the cotton zone,where didn't affected by frostbite,NDVI increased bigger, the canopy temperature was 26.4℃;where affected faintly by frostbite,the canopy temperature was 27.6℃;where affected seriously by frostbite,the canopy temperature was 29.3℃.Based on the difference of NDVI and canopy temperature, frostbite and the degree of the disaster can be monitored.
  • DONG He-Zhong, XIN Cheng-Song, LI Wei-Jiang, TANG Wei, ZHANG Dong-Mei, LUO Zhen
    Cotton Science. 2009, 21(4): 290-295.
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    Soil samples (0~20 cm) from 318 saline cotton fields randomly selected in the Yellow River Delta were used to study the [JP2]characteristics of salinity,contents of organic matter and major nutrients.The results showed that,of the total 318 saline fields tested,the lightly salty (LS) fields (salt content <2.5g·kg-1),moderately salty (MS) fields (2.5~4.49 g·kg-1) and severely salty(SS)fields (>4.50 g·kg-1) accounted for 44.3%,40.6% and 15.1%,respectively. The levels of N,P,K in MS fields were moderate,but K level in LS fields was low. The contents of organic matter,N and P were deficient,but K content was abundant in SS fields where cotton had been planted shorter than 5 years.In contrast,the contents of organic matter,N and P were moderate,but K content was relatively low in SS fields where cotton had been planted longer than 5 years.Regression analysis showed a significant linear correlation between soil salt content (y) and soil solution conductivity (x) at 25°С (y=3.4058x+ 0.1427,n=27,R2=0.9964**). Normal emergence and stand establishment rates were obtained when soil salinity was lower than 2 g·kg-1. Rates of emergence and stand establishment were reduced to 60%~78% and 45%~55%, respectively,when salinity was 2~3g·kg-1.The emergence and stand establishment rates were only about 40% and < 30%,respectively,in saline fields with salinity higher than 4 g·kg-1.Reasonable agronomic measures for stand establishment and chemical fertilization should be taken according to the characteristics of salinity and fertility,to improve cotton productivity in saline fields.
  • LIU Ling-Di, LI Cun-Dong, SUN Hong-Chun, LIU Lian-Tao, GAO Xue-Fei, FENG Li-Xiao
    Cotton Science. 2009, 21(4): 296-301.
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    The effects of water stress on cell damage, activities of protective enzymes and yield of different boll-weight genotypes in cotton were studied. The result showed that: under water stress, the content of MDA in these genotypes increased significantly according to both each genotype and each position and the increasing extent varied greatly with genotypes, stages and positions. Among the three species, the middle-boll genotype increased the least under water stress, the activities of SOD and POD increased at different position and for different genotypes in the most time, the activities of SOD in upper branch and middle branch leaves were the lowest at the 30-days age, and the changes were the same according to different position and for different genotypes, the change of the activity of SOD in upper branch and bottom branch leaves with middleboll genotype was more obvious than big-boll genotype and small-boll genotype. So far as the branch leaves of different locations at every period were concerned, the activity of POD in middle branch was the highest according to positions and stages, and the bottom branch was the second, and the upper branch was the lowest. The decreasing extent of boll weight and yield with middle-boll variety was the least which indicated that middle-boll genotype has higher drought tolerance ability than the other genotypes.
  • ZHOU Gui-Sheng, ZHOU Fu-Cai, XIE Yi-Ming, FENG Chao-Nian, YANG Yi-Zhong
    Cotton Science. 2009, 21(4): 302-306.
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    With Bt transgenic cotton plants of Kemian 1 as the testing material, temperature stress experiments (20℃, 25℃, 35℃,and 40℃) were carried out in temperature and light controlled compartments to investigate Bt insecticidal protein content in cotton leaves at flowering stage, the death rate of bollworm fed on cotton leaves, and the relationship between them. The results indicated that free amino acid content increased under temperature stress, with the biggest increment during the first 12 h. The content of soluble protein decreased, presenting a sharp decrement during the first 12 h. Bt insecticidal protein content in leaves subjected to temperature stress decreased significantly during the first 24 h, and the biggest decrements occurred during the first 12 h. The death rate of bollworm fed on the leaves subjected to 25℃ was higher than the death rates of bollworm eating the leaves subjected to 20℃ and 35℃. Under the same temperature, the longer the leaves were subjected to temperature stress, the lower the death rate of bollworm fed on these leaves was. The death rate of bollworm 72 h after eating the leaves was significantly higher than the death rate of bollworm 144 h after eating the leaves. Correlation analyses showed that the contents of Bt insecticidal protein, free amino acid, soluble protein and the death rate of bollworm 72 h after eating the leaves were significantly correlated at 0.05 level or at 0.01 level. The increment of free amino acid, the decrement of Bt insecticidal protein, and the death rate of bollworm 72 h after eating the treated leaves were significantly correlated at 0.05 level.
  • LI Shu-Na, WANG Shu-Xiang, WANG Zhan-Li, LI Hong-Ya, WANG Quan, ZHU Bao-Cheng
    Cotton Science. 2009, 21(4): 307-312.
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    ]In order to improve the spore production rate and spore quantity of antagonistic strain Bacillus subtilis LZ2-70 against Verticillium dahliae, the main factors influencing spore production were investigated using shake-flask fermentation method. Through single factor experiment and orthogonal experiment, the optimal shaking flask fermentation condition of strain Bacillus subtilis LZ2-70 was determined as follows: media composed of 0.5% Maltose, 2% soybean meal, 0.05% KH2PO4 and 0.1% MnSO4·H2O, initial pH 8.0, culture time 24 h and bottle filling capacity 30 mL/250 mL, 10% inoculums volume, fermentation time 36 h, fermentation temperature 30℃, rotating speed 200 r·min-1. Under this optimum condition, the spore quantity of strain Bacillus subtilis LZ2-70 achieves 6.6×109·mL-1 and the spore production rate achieves 98.94%.
  • SUN Feng-
    Cotton Science. 2009, 21(4): 313-318.
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    Effects of nitric Oxide and methylene blue on antioxidant capacity of the insect-resistant cotton seedlings roots under NaCl stress by determining activity of antioxidant enzymes and content of antioxidants were studied. The cotton seedlings were dealed with  nitric oxide and methylene blue(MB)respectively under normal and salt stress conditions,and enzyme activities of ascorbate peroxidase(APX),glutathione reductase(GR),dehydroascorbate reductase(DHAR), mono-dehydroascorbate reductase(MDAR) and content of ascorbic acid(ASA), glutathione(GSH/GSSG) were determined and analyzed. Results Showed that it promoted the growth of cotton seedlings by adding NO and significantly inhibited the growth of cotton seedlings by adding MB in normal conditions,lessened the inhibition of growth of cotton seedlings in salt stress by adding NO,and improved antioxidant capacity in roots. The activities of antioxidant enzymes and contents of antioxidants decreased by adding NO and NB at the same time in salt stress conditions,and the role of NO in the promotion of growth was removed.
  • BIAN Ru-Xin, LIU Ming, FAN Jun-Hua, CAO Juan, ZHANG Jian-Hua
    Cotton Science. 2009, 21(4): 319-323.
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    Sugar contents and cellulose deposition were monitored in the bolls of two varieties Xinhai 21 and Xinhai 28(YX-193 and XH-28) of island cotton in South Xinjiang in 2006 and 2007. After anthesis, total sugars in boll shells, seeds and fiber kept increasing and reached their maximum level at 21-28 days post anthesis (DPA). After that the sugar contents rapidly declined and reached the low level at the boll opening period. The cellulose was synthesized starting form 14 DPA and reached a peak at the boll opening stage. Soluble solids contents in fibers kept decreasing linearly during the boll development. Total soluble sugars, reducing sugars and their conversion rates showed an order as: fibers >seeds >boll shells, and were generally higher in YX-193 than in XH-28. The results indicate that sugar unloading and conversion capacities in bolls are related to fiber development and YX-193 has advantages over XH-28.
  • ZHOU Xiao-Feng, JIN Shuang-Xia, LI Xiang, LI Bao-Cheng, LIAN Wen-Ming, NING Xin-Zhu, LIU Li
    Cotton Science. 2009, 21(4): 324-329.
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    With three island cottons, Xinhai 17, Xinhai 14, 85H, shoot organogenesis and genetic transformation were studied in Xinjiang. The results were as follows: in vitro plant regeneration of meristem shoot tip is better, and the seedling rate of 90.5%was obtained. Frequency of inducing shoots reached maximum with 6-BA at 0.5 mg·L-1;  whereas number of budding shoots in per explant reached maximum with 6-BA at 1.5 mg·L-1; plantlet from shoot apical meristem repressed with 6-BA at 2.0 mg·L-1;The maximum budding shoots in per explant is four. Isolated meristem shoot tips infected with Agrobacterium tumefacien have strong endurance on infected damnification and kan, of which selective concentration reached 100 mg·L-1. The rate of plantlet from infected shoot tips on selection medium reached to 88.9%。
  • KUANG Meng, YANG Wei-Hua, XU Hong-Xia, WANG Yan-Qin, ZHOU Da-Yun, FENG Xin-Ai, WANG Jun-Fang
    Cotton Science. 2009, 21(4): 330-334.
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    There are three phases of DNA fingerprint technology based on molecular marker:the first is RFLP based on Southern blot,the second are several markers based on PCR and the third is SNP(single nucleotide polymorphism).Research and application of variety identification technology was discussed,four main molecular marker technologies applied in cotton variety identification were introduced in detail as follow:RFLP,RAPD,AFLP and SSR.Technology principle,research and application status,advantage and disadvantage about each molecular marker applied in cotton variety identification were expatiated.As compared,the viewpoint was put forward:SSR marker was the most appropriate molecular marker in cotton variety identification,and the necessity of construction of Chinese cotton DNA fingerprint database based on SSR marker technology was analyzed.
  • LIU Xin-Hu, ZHAO Xiao-Liang, WAN Chuan-Xing, ZHANG Li-Li
    Cotton Science. 2009, 21(4): 335-封三.
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    Effect of root exudates from the cotton material (CCRI 35) collected by water on cotton seedling growth and physiological activity was investigated. The results showed that root length, plant height, above-ground fresh weight,under-ground fresh weight of cotton seedling and cotton seed germination were promoted by root exudates at the low concentration, while restrained at high concentration.Root vigor,chlorophyll contents were reduced by root exudates at different concentrations. SOD activity was promoted by root exudates at the low concentration.