15 September 2009, Volume 21 Issue 5
    

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  • YE Wu-Wei, ZHAO Yun-Lei, WANG Jun-Juan, FAN Bao-Xiang
    Cotton Science. 2009, 21(5): 339-345.
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     A SSH library from the salt tolerant upland cotton cultivar, CCRI 35, was constructed using suppression subtractive hybridization (SSH) and pGEM-T Easy Vector under salt (0.4% of NaCl) stress at the first, third, fifth and seventh day after the beginning of salination. The library was evaluated to be of good quality with high efficiency of SSH. Total 200 positive clones were randomly selected and then sequenced, and 160 high quality sequences were obtained. According to BLAST screening and functional annotation, there were 62 unigenes (38.8%) without or only with weak similarities to the existent proteins in the NR database from NCBI and 42 unigenes(26.2%) significantly matched with hypothetical protein. While the unigenes assigned to stress-tolerance, metabolism, signaling conduction and transcription regulation accounted for 8.8%, 6.9%, 6.3% and 5.0%, respectively. The functions of these unigenes under salt stress were discussed.
  • ZHANG Dong-Dong, WANG Shi-Ying, GUO Xiao-Jun, JIANG Jun-Po, LIU Hui-Juan, ZHU Bao-Cheng
    Cotton Science. 2009, 21(5): 346-350.
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    In order to clone the gene which encodes the cotton Verticillium wilt antibacterial peptide A2, 15 amino acid residues of N-terminal of the antibacterial peptide A2 was inputted in NCBI to blast and the result showed a protein contained a fragment which was the same with the 15 amino acid residues of A2. Following the potein sequence, the gene of the protein was found and a pair of primers was designed according to the gene. A DNA fragment about 300 bp was obtained by PCR amplification using the genome of strain Bacillus amyloliquefaciens RS-25 as template. The PCR production was ligated into pET-28a plasmid by two restriction enzyme sites of BamHⅠand Hind Ⅲ. After colony PCR and restriction enzyme-digestion analysis, the PCR production was sequenced and sequencing analysis indicated that the PCR production was 354 bp and coded for a protein of 117 amino acid residues. This gene is named 25a2. The constructed expression vector pET-28a-25a2 was transformed into Eschrichia coli BL21 and colony PCR analysis was used to select the positive clones. The positive single clone was inoculated into LB medium and cultivated overnight with the kanamycin concentration about 100 mg·L-1. The cultivated bacteria was transfered into fresh medium with 10% inoculum and cultured at 37℃without antibiotics until the OD-600- reached 0.5~0.7. Then IPTG was added into the culture with the final concentration about 1 mmol·L-1 to induce the expression. Five hours later, the cultured cells were collected and ultrasonic crushed. SDS-PAGE analysis on expression product shows the molecular weight is about 15 kDa and coincides with the theory analysis.
  • WANG De-Long, YU Ji-Wen, YU Shu-Xun, DI Hong-Hong, FAN Shu-Li, SONG Mei-Zhen, ZHANG Jin-Fa
    Cotton Science. 2009, 21(5): 351-355.
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    SMART technique was used to construct the full length cDNA library of cotton seed. Total RNA was isolated from sea-island cotton embryo about 25~35 days. Full length ds-cDNA (Double-strained complement DNA) was amplified by using 21 round LD-PCR(Long-distance PCR). After SfiⅠdigestion and Chroma spin-400 fractionation, cDNAs (>500 bp) were ligated to pDNR-Lib vector and transformed to DH10B afterward. The total clones of the library was measured to be 5×106 with a titer of 1.5×108cfu·mL-1. The inserted fragments ranged from 0.5 kb to 2.5 kb, and the recombinant rate of the library was about 100%. 180 monoclones were selected to sequence, and 13 EST concerned with lipid metabolism were identified. These results show that the cDNA library can be used for screening genes of fatty acid  metabolism.
  • LI Lian, SHI Yu-Zhen, LI Jun-Wen, TANG Shu-Rong, LIU Ai-Ying, YU Ji-Wen, CHU Ping, YUAN You-Lu
    Cotton Science. 2009, 21(5): 356-360.
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    The stable BC2F4:6, BC2F4:7 lines, which were obtained from combination between 34B and Giza70 and backcrossed with 34B as recurrent parent, were used to analyse the correlation of fiber fineness related traits under three environments. The results showed:firstly, spinning uniformity index was significantly positive correlated with fiber length, uniformity and strength; significantly negative correlated with elongation. Fiber length was significantly positive correlated with strength, significantly negative correlated with elongation. And the correlation analysis indicated that it was possible to produce long, strong and good elongation fiber. Secondly, micronaire tested by HVI 900 was significantly negative correlated with strength and spinning uniformity index, maturity ratio was significantly positive correlated with micronaire, significantly negative with fiber linear density and maturity ratio were inconsistently correlated with other fiber traits under three environments. Fiber linear density was significantly positive correlated with spinning uniformity index, significantly negative correlated with length and strength, not significant with fiber uniformity, and negative with elongation. These indicated that linear density was more stable than micronaire and maturity ratio, so it is necessary to strengthen the study of linear density.
  • TANG Fei-Yu, CHENG Jin- , HUANG Wen-Xin- , MO Wang-Cheng, XIAO Wen-Jun
    Cotton Science. 2009, 21(5): 361-365.
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    Twelve characters within twenty high quality lines from the double crosses of twelve diverse upland cotton parents were measured in a randomized, complete block at the Agriculture Experimental Station of Jiangxi Agricultural University, Nanchang, China in 2007. The genetic variation, heritability, genotypic correlation and selection index of yield and fiber traits were calculated. The genetic variation coefficients of seed cotton yield per plant, lint yield per plant and bolls per plant were 16.64%, 14.71%, and 10.65%, respectively, higher than that of fiber traits varied from 1.39% to 6.72%. These results suggest a high availability of further improvement of yield in the population with high fiber quality, but of less selection potentiality for fiber traits. Among yield and yield components characters, lint percentage and boll weight produced the highest broad-sense heritability, 89.1% and 81.85%  respectively. The lowest broad-sense heritability was found in lint yield per plant(55.05%). Fiber traits all exhibited high broad-sense heritability, ranging from 79.07% to 89.47%. Lint yield per plant and bolls per plant were significantly and positively correlated, as were bolls per plant and number of fruiting branches. Twenty selection indices were constructed with one to four characters. The expected genetic advance of them were estimated. The efficiencies of the selection indices including the single plant lint yield itself and other highly correlated characters were better than those without lint yield per plant. The highest efficiency was recorded in the selection index consisting of lint yield per plant, bolls per plant, number of fruiting branch, number of boll position, higher than that of selection of single plant lint yield alone by 12.06%.
  • ZHU He-Qin, FENG Zi-Li, LIU Xue-Ying
    Cotton Science. 2009, 21(5): 366-370.
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    Widely growing of transgenic plant may be accompanied by ecological effects including changes in plant-associated with micro-flora. People pay more attention to environment risk assessment of transgenic plant. Most researches on Bt cotton have focused on invasiveness, gene flow to indigenous organisms, development of resistance in target pests, and  direct or indirect effects on non-target organisms and ecosystem.Some research have focused on the evaluation of   ecological effects of Bt cotton on microbial flora. But a few studies have been published to evaluate the ecological effects of CpTI cotton on microbial flora.Four CpTI cotton lines and their contrasts(non-CpTI cotton) were monitored for changes in total bacterial, actinomycetic and fungal population in 2006 and 2007.The results showed that the total bacterial, actinomycetic and fungal population of CpTI cotton and non-CpTI cotton changed in different growing period. The total bacterial population reached the maximum in flowering-bolling stage. So did the total actinomycetic and fungal population between budding stage and flowering-bolling stage. After that they declined gradually along with the decrepitude of cotton. The total bacterial population of CpTI cotton was significantly higher than that of non-CpTI cotton, especially in seedling stage and flowering-bolling stage. So did the total actinomycetic population of CpTI cotton from seedling stage to boll-opening stage.But the total fungal population was remarkably lower than that of non-CpTI cotton, especially in boll-opening stage.
  • LUO Hong-Hai, LI Jun-Hua, ZHANG Hong-Zhi, HE Zai-Ju, GOU Ling, ZHANG Wang-Feng
    Cotton Science. 2009, 21(5): 371-377.
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    Selected cotton cultivars of different genotypes as experimental materials,the effects of leaf-cutting and bud-thinning on photosynthetic rate,partitioning of 14C-assimilate and yield during flowering and boll-setting stage in high-yield cotton were studied.The results indicated that when leaves were cutted 1/4 at anthesis,the photosyntetic rate and the percentage 14C-assimilate of export and partitioning in boll of Xinluzao 13 at full bolling and boll opening stages increased.Conclusively,seed cotton yield and fiber quality had no obvious difference with those of control.However,the photosynthetic rate and photosynthesis products accumulation of aerial part of Biaoza A1 did not change when buds were thinned 1/4 at anthesis,but the speed of export and distribution percentage 14C-assimilates in boll increased,so the seed cotton yield exceeed that of control.These results suggest that it would be of great importance to regulate the relationship of source sink through the key cultivation techniques,according to different responses to the change of source and sink among varieties to obain super high yield and stable yield in Xinjiang.
  • QI Tong, HOU Zhen-An, YE Jun, RUAN Ming-Yan, 吕 Xin, HAO Zhi-Yong, SHI Li-Shang
    Cotton Science. 2009, 21(5): 378-382.
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    Effects of nitrogen rate and fertigation frequency on nitrogen uptake and yield of hybrid cotton were investigated through field experiment under drip irrigation conditions. The experiment comprised six treatments, with three nitrogen (N) rates (270, 360 and 450 kg·hm-2) and two fertigation frequencies (5 and 10 d). It was found that nitrogen rate and fertigation frequency significantly influenced nitrogen accumulation, dry mass and yield of hybrid cotton. Under 360 and 270 kg·hm-2 of N rates, fertigation frequency with 5 d promoted cotton growth and induced a significant increase in N accumulation and dry mass of hybrid cotton, but there were not statistically different between two fertigation frequency treatments for 450 kg·hm-2 of N level. It is recommended that increasing fertigation frequency could enhance yield of hybrid cotton in critical period of water and nutrition supply.
  • MA Li-Ying, CUI Jin-Jie, CHEN Hai-Yan
    Cotton Science. 2009, 21(5): 383-387.
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    The effects of transgenic Bt and Bt+CpTI cottons on soil enzyme activity in different stages were study under the greenhouse experiment condition. The results showed that from the seeding stage to the flower and boll stage, the activities of soil urease,polyphenoloxidase, acid phosphatase and alkaline phosphatase increased at first and then decreased. However, the activities of protease and the neutral phosphatase decreased at first and then increased.As for the same variety between the two neighboring stages of duration, the differences of the soil urease, protease, acid phosphatase and neutral phosphatase activities have reached the great remarkable level(P<0.01).Compared with nontransgenic cotton,the planting of transgenic cotton GK12 had a decrease of soil urease activity(23.06%) and a decrease of soil acid phosphatase activity(10.89%). The Bt+CpTI cotton sGK321 also decreased the enzyme activity. The transgenic Bt+CpTI cotton sGK321 caused a fewer disturbances on soil biological activities: compared with its parent nontransgenic cotton, it had an extremely significant effects on the activities of soil urease, protease, acid phosphatase and neutral phosphatase.
  • ZHAO Peng-Ju, WANG Deng-Wei, HUANG Chun-Yan, MA Qin-Jian
    Cotton Science. 2009, 21(5): 388-393.
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    Utilizing a quantum sensor and nonimaging hyperspectral spectrometer, 2 cotton cultivars 4 level densities planting, in north Xinjiang, multitemporal canopy hyperspectral data and photosynthetically active radiation data at cotton key growing stages were reoorded, and multivariate regression analysis method was used to analyze the correlated relationship between reflectance and cotton canopy FPAR, LAI, respectively. The results showed that the maximum correlation coefficients between hyperspectral data and FPAR, LAI occurred at 813 nm, 758 nm wavelength,respectively;based on the linear regression equation between reflectance spectrumρ813 and ρ758 and FPAR, LAI, respectively, their correlation coefficients were significant between measured FPAR and estimated FPAR, measured LAI and estimated LAI (rFPAR =0.7199**,rLAI =0.6430**,α=1%, n=70). The regression function accuracies were 96.5%,81.7%, respectively.The maximum value of correlation coefficient between the first derivative spectral data and LAI occurred at 734 nm wavelength,but the correlation coefficient was not significant between the first derivative spectral data and FPAR from 350 nm to  2500 nm wavelength. According to the first derivative spectral value ρ′734 estimate for LAI, measured LAI and estimated LAI was significantly relevant (rLAI=0.6947**,α=1%, n=70),the regression function accuracy was 82.4% and was nearly to the prediction precision of the LAI regression modeling at 758 nm wavelength reflectance. The study showed that canopy hyperspectral data can be used for real-time, nondestructive and quantitative estimate of FPAR, LAI.
  • WANG Wei
    Cotton Science. 2009, 21(5): 394-398.
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     A material when a foreign chromosome is replaced into the background parent chromosome is called substitution line. Whereas, a material chromosome segments are replaced, is called introgression line. F2 and F2:3 populations of (TM-1×IL6) were constructed with the background parent TM-1 and introgression line 6(IL6) of chromosome 6 of island cotton 3-79. We investigated 18 important agronomic traits in two years' field experiment and made a genetic evaluation. Besides, molecular data of F2 population were obtained and QTL mapping and single marker analysis were carried out. Segments introgressed into IL6 included chromosome 6, non-chromosome 6 of 3-79 and most TM-1. Yield and quality traits of IL6 parent were more excellent than TM-1. Likewise, a genetic analysis was made and alleles increasing genetic effects from IL6 were found. It means that segments introgressed into IL6 play a crucial part. Agronomic traits and genetic analysis indicated that there were major genes controlling total bolls at full-growing stage, boll numbers per plant, lint percent, fruit branches per plant, fiber strength and the micronaire in the segments. QTL mapping and single marker analysis showed that there were QTLs controlling boll numbers per plant, lint percent, fruit branches per plant, plant height, seed index and uniformity ratio in the segments.
  • WU Da-Peng, FANG Wei-Ping, JI Dao-Fan, ZHU Shui-Jin
    Cotton Science. 2009, 21(5): 399-404.
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    Two cultivars of Gossypium barbadense and five upland cotton (G. hirsutum) cultivars were used to cross with CCRI 12, and 14 hybrids and its F1 and F2 populations were produced. The inheritance and its utilization of CCRI 12 in breeding program for Verticillium wilt resistance were studied in this experiment, using the method of seedling inoculation. The results showed that, for the combinations used CCRI 12 as paternal parent, the ratio of the resistant and susceptible seedlings in F2 were done according with the 3:1 separation law, which indicated that the Verticillium wilt resistance in G.barbadense was a qualitatively inherited trait controlled by singly dominant gene, as well as the tolerant trait in upland cotton cultivars. However, for those using CCRI 12 as female parent, the frequency of the resistant seedlings was much higher than those using CCRI 12 as paternal parent, and more than 90% of seedlings in the F2 populations were disease resistant ones, which was out of the separation law greatly. It indicated that there were some disease resistant substances in the cytoplasm in CCRI 12, the inheritance of the Verticillium wilt resistance in CCRI 12 was like that of cytoplasm genetic characters. The present research work indicated that the parent selection and cross method were very important in Verticillium wilt resistant cotton breeding program, and CCRI 12 is a very special germplasm in cotton breeding for Verticillium wilt resistance.
  • ZHOU Da-Yun, YANG Wei-Hua, XU Hong-Xia, WANG Yan-Qin, FENG Xin-Ai, KUANG Meng, HOU Ai-Ling
    Cotton Science. 2009, 21(5): 405-409.
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    ]In order to know the effect of temperature on germination and dormancy of cottonseeds during storage, fuzzy and coated cottonseeds with dormancy were studied, germination,germination potential(GP), the percentage of fresh seeds(PF), fresh weight and length of seedling were analyzed and compared during and after 4 years storage. Fuzzy and coated samples with dormancy were stored at room temperature, 0℃and -18℃ from April 2005 to April 2008. Seeds containing 11% or less moisture were sealed in the plastic bags and stored. One hundred seeds were sampled from each sample to conduct germination trials at 30℃ for a 7 days period every year. Effects of one factor of temperature,and two factors reciprocal action of  temperature and storage periods were studied during 4 years storage. The one factor results showed that temperature was not significant to the change of germination and GP of coated seeds and germination of fuzzy seeds, it was obviously significant to GP of fuzzy seeds and  PF of both fuzzy and coated. The reciprocal action effects of temperature and years effected the germination and dormancy of fuzzy and coated seeds significantly. Effects of temperature on fresh weight and length of seedlings were different between fuzzy and coated seeds. Lower temperature could postpone the dormancy breaking, decelerated the speed of the increase of germination, put off the deterioration, and increased the longevity of cottonseeds.The results of 0℃ were better than the other two temperture.
  • ZHANG Xiao-Quan, WANG Xue-De, JIANG Pei-Dong, ZHU Wei
    Cotton Science. 2009, 21(5): 410-414.
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    In order to utilize heterosis of interspecific hybrid cotton between Gossypium barbadense and G. hirsutum and increase its hybrid seed yield, 10 three-way cross hybrids and 4 single cross hybrids were produced using cytoplasmic male sterile, maintainer and restorer lines in G. barbadense and G. hirsutum, and were evaluated in a Randomized Complete Block Design with three replications during 2005-2006 at Zhejiang University Farm. The results showed that the cotton yield of three-way cross hybrids was very significant higher than that of single cross hybrids in G. barbadense, and similar with single cross hybrids between G. barbadense and G. hirsutum, but lower than single cross hybrids in G. hirsutum. For the cotton fiber quality, three-way cross hybrids were not only better than single cross hybrids in G. hirsutum but also had a same level to single cross hybrids between G. barbadense and G. hirsutum. Especially, three-way crossing was able to produce higher hybrid seed yield than single crossing in G. barbadense although was not in G. hirsutum. It is suggested that early mature sterile lines in G. barbadense, high lint percentage and boll weight maintainer and restorer lines in G. hirsutum, and with relative uniformity in plant shape using as parents might be considered in the breeding program of three way cross hybrid.
  • TANG Wen-Wu, HUANG Ying-Jin, WU Xiu-Lan, XIAO Wen-Jun
    Cotton Science. 2009, 21(5): 415-419.
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    Combining ability and genetic effects of yield and fiber quality characters were investigated by way of 10×6 NCII cross design with 10 super fiber varieties and 6 insect-resistant transgenic varieties. The results showed that, Bt gene was dominant inheritance in F1 hybrids. A8 showed the best general combining ability (GCA) in yield characters, A4 expressed the best GCA in fiber quality characters. Combinations of A7×B5, A6×B1 expressed higher specific combining ability(SCA). But the GCA of parents and SCA of combination have no significant correlation. Genetic effects analysis showed that the additive variation was principal in boll weight (BW), lint percentage (LP), span length (SL), fiber strength (FS), elongation rate (ER). While seed cotton yield (SCY) and lint yield (LY) mainly by their interactions. SL, BW and FS had high narrow heritability (HN2), while SCY and LY were had low HN2.
  • MIAO Pei-Ming, FAN Ling, SHI Wei-Jun, WU Maierjiang, XU Li-Min, MA Jun
    Cotton Science. 2009, 21(5): 420-426.
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    The genetic diversity among 65 lines of cotton germplasm was analyzed by TRAP (Target Region Amplification Polymorphism) markers. From 100 pair of primers, 10 pairs of polymorphic primers were selected. A total of 252 bands were amplified from 10 pairs of primers by TRAP-PCR, of which 210 bands (about 83.33%) were polymorphic. The 65 lines of cotton germplasm were clustered into six groups at GS (genetic similarities) = 0.8431 and the link map was set up by UPMGA method in NTSYSpc 2.1(numerical taxonomy and multivariate analysis system).The results showed that the germplasm clusters had certain relationship with the germplasm origins and the morphologic characters. The results suggested that the new type of molecular maker method, TRAP, was suitable for genetic identification, cluster of cotton germplasm.
  • LUO Xin-Ning, CHEN Bing, ZHANG Ju-Song, JIANG Ping-An, LOU Shan-Wei, PENG Xiao-Feng, HE Jia-Lin
    Cotton Science. 2009, 21(5): 427-430.
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    Based on the cotton pot experiment involving four varieties (three were upland cotton and one was sea island cotton) and two N treatments and field experiment with different N treatments,the spatial differences of N and Chlorophyll contents (SPAD value) in the four upper fully expanded leaves and their relationships with whole leaf and plant N concentration were investigated. The results showed that leaf N contents and SPAD values at different leaf positions differed obviously,leaf N contents and SPAD values increased and differences among different position leaves decreased at high N rates. The sensitivity of the fourth leaf from the top to N was the highest,and next was that of the second leaf from the top was the lowest,while that of the first and the third leaves varied within the varieties. The SPAD value of the fourth leaf from the top was most correlated with whole leaf and plant N concentration among the top four leaves through the bud stage,initialflowering stage. The CV of SPAD value of the fourth leaf from the top was the smallest under optimal N level. It was suggested that the fourth leaf from the top was the most ideal indicator when using SPAD value or leaf color difference to diagnose N status in cotton.
  • LIU Wei-Yang, ZHENG De-Ming, JIANG Yi-Juan, FAN Ye-Kuan
    Cotton Science. 2009, 21(5): 431-封三.
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    The systematic study was employed to study the growth characteristics and the accumulation law of N, P, K nutrients. Three treatments were designed in cotton fields including low-yield, medium-yield, high-yield cottons. Results showed that the stem length of high-yield cotton had no such changes with medium-yield and low-yield cottons,but the stem increase of high-yield cotton stopped before medium and low yield cottons. The numbers of total bolls for medium and low yield cottons were much lower than the highyield, and the spatial distribution of bolls for the high-yield cotton was more reasonable. The accumulations of N, P2O5, K2O in cotton plant could be described by the Logistic curve equation. Compared to the medium and low yield cottons, the fast increase period and the sustained time of the high-yield cotton were obviously discrepant. The total accumulations of Nitrogen, phosphorous and potassium for the high-yield cotton were 515.3, 126.4, 591.9 kg·hm-2, respectively. Compared to the medium and low yield cottons, the nutrient coordination of the high-yield cotton was better, and the total uptake nutrient was more. These showed that it is possible to achieve largescale high-yield cotton by reasonable application of cultural factors in the South Xinjiang cotton district.