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Cotton Science
Volume 34 Issue 3
15 May 2022
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    Functional analysis of GhEIN3 gene in response to cotton Fusarium wilt stress
    Zhao Zengqiang,Zhang Xi,Li Xiaoling,Zhang Wei
    2022, 34(3):  173-186.  doi:10.11963/cs20210046
    Abstract ( 38 )   HTML ( 168)   PDF (16700KB) ( 20 )  
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    [Objective] Ethylene (ET) insensitivity protein 3 (EIN3)/EIN3-like (EIL) family genes are key genes in ethylene pathway, which are involved in the biological stress response of plants. Its function was studied to provide a basis for analyzing the response mechanism of upland cotton to Fusarium wilt. [Method] GhEIN3 of EIN3/EIL gene family were selected from TM-1 genome database based on the data of gene expression profile from root infected with cotton Fusarium wilt in our laboratory. The gene coding protein and related sequence information were analyzed by bioinformatics methods, and the expression pattern of GhEIN3 in Fusarium wilt and different hormone treatments were analyzed by real-time fluorescent quantitative polymerase chain reaction (qRT-PCR). The role of GhEIN3 in disease resistance was preliminarily analyzed by virus induced gene silencing (VIGS) technique. [Result] A new gene GhEIN3 was isolated from upland cotton (Genbank accession: KY744279), which was from EIN3/EIL family. Bioinformatic analysis showed that the open reading frame of this gene was 1 842 base pairs. The encoded amino acids sequence contains a typical EIN3 domain. After Fusarium oxysporum f. sp. vasinfectum (FOV) and different exogenous hormones ET, salicylic acid (SA) and jasmonic acid (JA) treatment, the expression of GhEIN3 was up-regulated by FOV and ET induction, but down-regulated by SA and JA induction. The VIGS results showed that the GhEIN3 gene silenced plants were more susceptible to FOV than the control. The results of gene expression analysis showed that compared with the control, the expression levels of pathogenesis-related genes PR1, PR2, and PR4, ethylene response factor gene ERF1 and 1-aminocyclopropane-1-carboxylate oxidase gene ACO were decreased, while the expression level of PR5 gene was increased after GhEIN3 gene silencing. [Conclusion] A EIN3/EIL family gene GhEIN3 identified from G. hirsutum responds to Fusarium wilt and hormones ET, SA and JA induction. Combined the GhEIN3 gene expression analysis of the cotton plants under ET, SA, JA and FOV treatments with VIGS results, this study showed that the GhEIN3 gene plays a positive role in the process of cotton to resist Fusarium wilt.

    Cloning and disease-resistant function of a senescence-associated gene GhSAG101 in Gossypium hirsutum
    Wu Jianfeng,Fan Zhihao,Wu Lianjie,Hu Xiaowang,Han Zhili,Gao Wei,Long Lu
    2022, 34(3):  187-197.  doi:10.11963/cs20210030
    Abstract ( 30 )   HTML ( 35)   PDF (11627KB) ( 14 )  
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    [Objective] Screening and study of cotton disease resistance genes will provide theoretical basis and genetic resources for cotton disease resistance breeding. [Method] The senescence associated gene 101 (GhSAG101) was cloned from Gossypium hirsutum L. TM-1. The expression levels of GhSAG101 in different organs of cotton seedlings, as well as in cotton roots under Verticillium dahliae inoculation were analyzed by real time quantitative polymerase chain reaction (qRT-PCR). The conserved domains and active sites of GhSAG101 were predicted through protein sequence analysis. Knockdown lines of GbSAG101 generated through virus induced gene silencing (VIGS) system were employed for the examination of Verticillium dahliae resistance by inoculation of the whole plant and detached leaves, and the empty vector transferred plants were used as the negative control. [Result] The coding region of GhSAG101 gene is 1 764 bp in length, encodes a protein with 587 amino acid residues that includes a nucleophilic elbow and the conserved domains belonging to the abhydrolase superfamily and enhanced disease susceptibility 1 (EDS1) family. The highest expression level of GhSAG101 was found in roots, followed by stems, and the lowest expression level was found in leaves. GhSAG101 expression was induced by V. dahliae infection in roots at the middle and late infection stage. Suppression of GhSAG101 expression in cotton reduced the spread of V. dahliae and retarded the disease process. [Conclusion] GhSAG101 negatively regulates cotton resistance to V. dahliae and can be used as a candidate gene in the breeding of cotton with improved disease resistance.

    Effects of increasing application depth and decreasing nitrogen rate on dry matter, nitrogen accumulation and distribution, and yield of direct seeding cotton after rape harvest
    Li Fei,Guo Lili,Zhao Ruiyuan,Yin Lingjie,Wang Jiazhen,Li Caihong,He Shujun,Mei Zhengding
    2022, 34(3):  198-214.  doi:10.11963/cs20210072
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    [Objective] The effects of nitrogen application depth and amount on cotton dry matter and nitrogen accumulation, distribution and yield in rape-cotton rotation cropping system around Dongting Lake cotton planting region were studied, so as to provide a scientific basis for reducing the application and increasing the efficiency of chemical fertilizer of direct seeding cotton after harvesting rape. [Method] The nitrogen application experiment was carried out at Changde, Hunan Province in 2018 and 2019. A field experiment was conducted using Xiang FZ001 with the treatments of two nitrogen application depths (15 cm and 5 cm, marked by H15 and H5), and five nitrogen rates (0, 90, 180, 270 and 360 kg·hm-2, marked by N0, N90, N180, N270, N360, respectively). [Result] The trends of cotton nitrogen and dry matter accumulation were consistent, and the accumulation dynamics were in line with the logistic model. Deep application of nitrogen fertilizer increased the accumulation of nitrogen and dry matter in cotton plants and their distribution proportion in reproductive organs. With the increase of nitrogen application rate, the accumulation of nitrogen and dry matter and their distribution proportion in reproductive organs all showed a trend of increase at first and decrease afterward. Under the interaction of nitrogen application depth and amount, the accumulation of nitrogen was maximum under H15N270; the proportion of nitrogen distribution in reproductive organs, dry matter accumulation and its distribution proportion in reproductive organs were maximum under H15N180. From the characteristic values of nitrogen and dry matter accumulation, it could be seen that the accumulation and distribution of nitrogen and dry matter in cotton was basically sychronized, with the average rapid accumulation of nitrogen starting slightly earlier than that of dry matter accumulation, indicating that cotton nitrogen absorption was likely the basis and premise of dry matter accumulation. Compared with H5, H15 improved the utilization efficiency of nitrogen fertilizer, and the apparent utilization rate, agronomic utilization rate and biased productivity of nitrogen fertilizer decreased with the increase of nitrogen application rate. With the interaction of nitrogen application depth and amount, the H15N180 treatment could obtain desirable apparent utilization rate, agronomic utilization rate, biased productivity and production efficiency and highest seed cotton yield. [Conclusion] This study indicated that applying suitable nitrogen application rate and increasing fertilization depth could promote high yield. And it was recommended that the depth of nitrogen application should be increased from 5 cm to 15 cm,and the quantity should be reduced from 270-300 kg·hm-2 to 180 kg·hm-2 for the tested region.

    Identification and functional analysis of the ACX gene family in Gossypium hirsutum L.
    Tian Yibo,Pan Ao,Chen Jin,Zhou Zhonghua,Yuan Xiaoling,Liu Zhi
    2022, 34(3):  215-226.  doi:10.11963/cs20220012
    Abstract ( 35 )   HTML ( 35)   PDF (18092KB) ( 36 )  
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    [Objective] The acyl-coenzyme A oxidase (ACX) gene family was identified in Gossypium hirsutum and their expression patterns were analyzed, which would lay the foundation for further studies of the function of ACX genes. [Method] The members of ACX gene family in G. hirsutum genome were identified by bioinformatics method, and their physical and chemical properties, gene structure, evolutionary relationships, gene replication, cis-acting elements in promoter region and expression patterns were analyzed systematically, and the virus-induced gene silencing (VIGS) technique was used to preliminary explore the function of GhACX16 gene. [Results] A total of 20 ACX genes were identified in G. hirsutum genome and distributed on 13 chromosomes, which were divided into 4 subfamilies by cluster analysis. The results of non-synonymous mutation rate/synonymous mutation rate (Ka/Ks) analysis showed that the ACX genes in G. hirsutum had undergone strong purifying selection. The promoter region of ACX genes contain cis-acting elements related to heat stress, drought, plant hormone responses, etc. The expression patterns under abiotic stresses showed that the ACX genes responded obviously to high-temperature, low-temperature, salt, and simulated drought stress. The expression patterns of ACX genes in the anthers at different developmental stages of high temperature-tolerant/sensitive cotton lines under high temperature conditions were analyzed, and it was found that the expression levels of GhACX5 and GhACX16 changed obviously. Compared with the negative control, the GhACX16 silenced cotton seedlings showed obvious characteristics of high temperature tolerance, and the proline content, chlorophyll content, and catalase activity were significantly higher, while the malondialdehyde content was significantly lower in the leaves. [Conclusion] The identification and expression patterns of ACX genes in G. hirsutum showed that ACX genes were widely involved in abiotic stresses response, and GhACX16 gene was preliminary speculated to play an important role in high temperature stress response according to the VIGS verification and physiological and biochemical analysis.

    Electrophysiological and olfactory responses of Pectinophora gossypiella adults to extracts from four kinds of Chinese medicine plants
    Yin Haichen,Xu Dong,Wu Huaiheng,Cong Shengbo,Yang Tiantian,Wan Peng
    2022, 34(3):  227-234.  doi:10.11963/cs20210043
    Abstract ( 31 )   HTML ( 9)   PDF (1332KB) ( 13 )  
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    [Objective] The purpose of this study is to utilize Chinese herbal medicine resources and to establish new monitoring and control technology of Pectinophora gossypiella. [Method] The electrophysiological and olfactory responses of P. gossypiella to the extracts of Zanthoxylum bungeanum, Cortex phellodendri, Saussurea costus, and Rhus chinensis were studied in this study, and the synergism of the extracts was further studied. [Result] The extracts of Z. bungeanum and S. costus could induce strong electrophysiological responses of P. gossypiella when the concentration of odor source was 15 g·L-1. The results of olfactory test showed that the extracts of Z. bungeanum had significant attraction to both female and male adults, and the selected response rates were 67.26% and 76.72%, respectively. The extract of S. costus had a very significant attraction to male adults, and the selected response rate was 71.19%. Further studies showed that the attractive effect of these two extracts was similar to that of cotton plants. Z. bungeanum extract mixed with linalool with the ratio of 3:7 and 6:4 (by volume), showed significantly synergistic effect on male and female insects respectively. [Conclusion] Based on a series of indoor experiments, two potential attractant, the mix ratio of Z. bungeanum extract to linalool was 3:7 and 6:4, with high activity to P. gossypiella were screened, which can provide support for the detection and green control of P. gossypiella.

    Analysis of variation of cotton leaf temperature and its influencing factors based on infrared sensors
    Wang Yaru,Yang Beifang,Lei Yaping,Xiong Shiwu,Han Yingchun,Wang Zhanbiao,Feng Lu,Li Xiaofei,Xing Fangfang,Xin Minghua,Wu Fengqi,Chen Jiale,Li Yabing
    2022, 34(3):  235-246.  doi:10.11963/cs20210064
    Abstract ( 35 )   HTML ( 18)   PDF (5537KB) ( 8 )  
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    [Objective] Leaf blade is a plant organ that is sensitive to environmental changes. Leaf temperature is an important physiological index of plants. This study aims to explore the day-night variation characteristics of leaf temperature and to clarify the influence of meteorological factors on leaf temperature. [Method] In this study, the temperature of cotton leaves was automatically monitored in real-time based on infrared temperature sensors, and then the characteristics of day-night variation of cotton leaves temperature in different growth periods and under different weather conditions were explored, and the relationship of meteorological factors with leaf temperature and leaf-air temperature difference were explored by correlation analysis, stepwise regression analysis and path statistical analysis. [Result] The day-night variation of leaf temperature was different in various weather conditions and growth stages, and the variation range of leaf temperature was less than that of air temperature. Environmental factors were significantly correlated with cotton leaf temperature (except precipitation) and leaf-air temperature difference (except water vapor pressure) (P<0.05). The correlation between air temperature and leaf temperature was the highest (r = 0.890). The correlation between air relative humidity and leaf-air temperature difference was the highest (r = 0.825). The results of path analysis showed that the order of factors’ effect on leaf temperature according to decision coefficient was air temperature > photosynthetic active radiation > water vapor pressure. Photosynthetic active radiation and water vapor pressure mainly indirectly affected the change of leaf temperature through air temperature. The order of the effects of analyzed environmental factors on the leaf-air temperature difference according to the decision coefficient is air relative humidity > sunshine time > water vapor pressure. Sunshine time and water vapor pressure indirectly affect the change of leaf-air temperature difference through air relative humidity. [Conclusion] This study explored the day-night dynamic changes of cotton leaf temperature, and preliminarily explored the comprehensive effects of environmental factors on leaf temperature and leaf-air temperature difference. The results provide reference for cotton production and intelligent management.

    Effects of chemical topping on dry matter accumulation and distribution of cotton in Southern Xinjiang
    Hu Yukai,Zhao Shuzhen,Dong Hongqiang,Wei Yonghai,Tian Yugang,Chen Jialin,Dong Helin,Ma Xiaoyan,Feng Lu,Zhai Yunlong,Chen Guodong
    2022, 34(3):  247-255.  doi:10.11963/cs20210048
    Abstract ( 38 )   HTML ( 27)   PDF (1895KB) ( 10 )  
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    [Objective] This study aims to analyze the effects of chemical topping on cotton dry matter accumulation and distribution. [Method] Three upland cotton varieties (Xinluzhong 37, Xinluzao 77, and Xinluzhong 82) were selected as the experimental materials. A two-factor split plot design was adopted, taking four topping treatment (75, 105, 135 g·hm-2 mepiquat chloride, and manual topping) in the main plot, three cotton varieties in the secondary plot, to study the effects of the chemical topping on dry matter accumulation and distribution of cotton in the arid area of Southern Xinjiang. [Result] The 75 g·hm-2 mepiquat chloride treated plants had the lowest total dry weight of vegetative organs at 7-28 days after the treatment, which was 2.91%-31.12% lower than that of 105 g·hm-2, 135 g·hm-2 treatment and manual topping. The dry matter accumulation of reproductive organs of 75 g·hm-2 and 105 g·hm-2 chemical topping treatments were 9.18% and 13.93% higher than that of manual topping at 42 days after treatment, respectively. Under 75 g·hm-2 treatment, the yield of Xinluzhong 37, Xinluzao 77, and Xinluzhong 82 was 8.64%, 4.54% and 18.56% higher than that of manual topping, respectively. [Conclusion] Applying of 75 g·hm-2 mepiquat chloride had more obvious inhibition effect on dry matter mass of vegetative organs and could effectively increase cotton yield. There was significant or extremely significant interaction between chemical topping dosage and cotton varieties on dry matter accumulation in cotton vegetative organs.

    Effects of arbuscular mycorrhizal fungi on photosynthetic characteristics and mesophyll cell ultrastructure of cotton under arsenic stress
    Gong Minggui,Liu Kaiyang,Wei Yanan,Bai Na,Qiu Zhijun,Zhang Qiaoming
    2022, 34(3):  256-266.  doi:10.11963/cs20220022
    Abstract ( 33 )   HTML ( 13)   PDF (6348KB) ( 32 )  
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    [Objective] The effect of arbuscular mycorrhizal fungi (AMF) inoculation on photosynthetic characteristics and mesophyll cell ultrastructure in cotton under arsenic stress were studied. [Method] Cotton germplasm Dalingmian 69 were used as the test material in the pot experiment. Two species of AMF including Rhizophagus intraradices and Funneliformis mosseae were inoculated into the cotton roots respectively in order to investigate the effects of AMF on stomatal morphological characteristics, gas exchange parameters, chlorophyll fluorescence parameters and mesophyll cell ultrastructure under different arsenic concentrations (0, 100, 200 mg·kg-1) in soil. [Result] (1) Arsenic stress significantly reduced stomatal length, width, density, conductance, transpiration rate, intercellular CO2 concentration and net photosynthetic rate, and significantly affected chlorophyll fluorescence parameters of cotton leaves. However, compared with the control treatment, inoculation of AMF increased the length, width, aperture, density, and conductance of cotton stomata, and significantly increased the net photosynthetic rate, transpiration rate, intercellular CO2 concentration, actual photosynthetic efficiency, the maximum photochemical quantum yield and photochemical quenching coefficient, and reduced the non-photochemical quenching coefficient under different concentrations of arsenic. Under 100 and 200 mg·kg-1 arsenic concentration, the net photosynthetic rate and actual photosynthetic efficiency of F. mosseae treatment were significantly higher than those of R. intraradices treatment. (2) Arsenic stress led to the gradual swelling of chloroplast, the overall structure was destroyed, the disordered and uneven arrangement of grana thylakoids, the loose and deformed arrangement of cristae in mitochondria, the appearance of voids in some mitochondrial structures, and the fuzzy double-layer membrane structure. However, under arsenic stress, the lamellae of chloroplast grana in the mesophyll cells of cotton inoculated with AMF were clearly visible, and the cristae in mitochondria were closely arranged, and the degree of arsenic damage was significantly reduced. The palliative effect of F. mosseae was even stronger. [Conclusion] Under arsenic stress, inoculation of F. mosseae and R. intraradices significantly increased the net photosynthetic rate and light energy utilization efficiency, and improved the photosynthetic capacity of cotton, and reduced the damage of arsenic to the ultrastructure of mesophyll cells. And the inoculation effect of F. mosseae was better.