棉花品种遗传纯度的SSR分子标记鉴定技术研究

为建立适合于SSR标记的棉花品种纯度鉴定方法,利用78个SSR标记对12个棉花常规品种进行标记基因型分析.通过比较不同品种不同单株标记基因型,发现棉花品种的非纯SSR位点存在3种主要类型.通过综合分析非纯SSR位点率和异型单株率对品种遗传纯度的影响,建立了利用SSR标记在分子水平上鉴定棉花品种纯度的方法.利用该方法鉴定的12个棉花品种中有3个品种(C5、C9和Cll)的遗传纯度在98％以上,非纯SSR位点和异型株均较高的2个品种(C3和C6)遗传纯度分别为67.31％和31.79％,该方法弥补了以往分子纯度计算方法中只考虑单株混杂、不考虑SSR位点混杂的缺陷,可比较客观地反映品种的遗传纯度状况.     

The Effectiveness of HA-GGE Biplot Application in Analyzing the Data from the National Cotton Cultivar Regional Trial in the Yangtze River Valley

A large number of regional crop trials have demonstrated the ubiquitous existence of genotype × environment interactions (G×E), which make it complicated to select superior cultivars and identify the ideal testing sites. The GGE (genotype main effect plus genotype × environment interaction) biplot is the most powerful statistical and graphical displaying tool available for regional crop trial dataset analysis. The objective of the present study was to demonstrate the effectiveness of the biplot in evaluating the high and stable yields of candidate cultivars simultaneously, and in delineating the most adaptive planting region, analyzing trial location discrimination ability and representativeness, and identifying the ideal cultivar and trial locations. The lint cotton yield dataset with nine experimental genotypes and 17 test locations (three replicates in each) was collected from the national cotton regional trial in the Yangtze River Valley (YaRV) in 2012. The results showed that: (1) the effects of genotype (G), environment (E), and genotype × environment interaction (G×E) were significant (P < 0.01) for lint cotton yield. Differences among environments accounted for 78.7% of the treatment total variation in the sum of squares, whereas the genotype main effect accounted for 8.7%, and the genotype × environment interaction accounted for 12.6%. (2) The “ideal cultivar” and “ideal location” view of the HA-GGE biplot identified Zhongcj408 (G2) and Nannon12 (G9) as the best ideal genotypes; Cixi in Zhejiang Province and Jiangling in Hubei Province were the most ideal locations.(3) The “which-won-where” view of the biplot outlined the adaptive planting region for each experimental cultivar. (4) The “similarity among locations” view clustered the trial locations into four groups, among of which the two outlier locations, Shehong (SH) and Chengdu (QBJ), located in Shichuan Basin in the upper reaches of YaRV, were clustered in one group, whereas the Nanyang (NY) of Henan Province at the northern edge of YaRV was singled out as a sole group. Such location clustering results implied an apparent association with the geographical environment.     

Leaf Temperature and Transpiration of Field Grown Cotton and Soybean under Arid and Humid Conditions

Abstract

Abstract : Cotton (Gossypium hirsutum L.) and soybean (Glycine max (L.) Merr.) cultivars were grown under arid (Urumqi, Xinjiang, China) and humid (Matsudo, Chiba, Japan) conditions to analyze their abilities to adapt to arid conditions in terms of transpiration, leaf movement and leaf temperature. Under the arid condition, the leaf temperature of the cotton cultivars was higher than that of the soybean cultivar and the air temperature. There was no significant difference in leaf temperature among the cotton and soybean cultivars under the humid condition. The flow rate of stem sap in the cotton cultivars under the arid condition was always higher than that in the soybean cultivar, and was largely affected by vapor pressure deficit (VPD). Under the humid condition, however, the flow rates of stem sap were lower in the cotton cultivars than in the soybean cultivars. These results indicate that cotton can avoid heat stress by the high transpiring ability possibly supported by well-developed root systems, which leads to higher drought resistance under the arid condition. Soybean would adapt to arid conditions by the combination of paraheliotropic leaf movement and reduced transpiration.     

Potassium distribution in root and non‐root zones of two cotton genotypes and its accumulation in their organs as affected by drought and potassium stress conditions

A rhizobox experiment was conducted to compare the differences of soil potassium (K) distribution and absorption between two cotton (Gossypium hirsutum L.) genotypes under drought and K‐deficit conditions. Treatments included two levels of water (drought and optimum soil moisture: 25% and 35% volumetric water content) and K fertilizer rates (0 and 0.48 g potash kg^?1 soil) applied to two cotton genotypes (namely HEG and LEG). Both the genotypes showed significant differences in total K accumulation without exogenous K addition. After absorption, soil content of the readily available potassium (RAK) decreased rapidly. This promoted the conversion of the mineral K into slowly available potassium (SAK). Drought significantly decreased the cotton growth and K use efficiency, and thereby reduced the effect of K fertilizer. Consequantly, the contents of RAK and SAK were greatly increased. However, K bioavailability was decreased under water stress conditions. Differences in root parameters and soil microorganisms between two cotton genotypes were significantly increased and had marked relations with available soil K contents. This study provides important information for understanding the mechanism of K use efficiency, especially under water and K stress.     

通过GbF3’H基因单独沉默及其与GbCHI和GbDFR基因共沉默研究其在海岛棉中抗枯萎病功能

【目的】通过沉默海岛棉GbF3’H基因及共沉默GbF3’H、GbCHI和Gb DFR基因,研究其在海岛棉抗枯萎病中的作用。【方法】以海岛棉抗病材料06-146为研究对象,GhCLA1基因为阳性对照,空载体为阴性对照,构建海岛棉TRV2-Gb F3’H沉默载体,协同课题组前期构建的TRV2-CHI和TRV2-DFR载体,利用病毒诱导的基因沉默技术(Virus-induced gene silencing,VIGS)分别进行Gb F3’H基因单独沉默以及GbF3’H、GbCHI和Gb DFR这3种基因共沉默试验。通过实时荧光定量聚合酶链式反应(Quantitative real time-polymerase chain reaction,q RT-PCR)分析各处理样品中基因沉默情况;设置室内接种枯萎病菌试验测定病情指数,分析各沉默材料对枯萎病的抗性差异。【结果】q RT-PCR结果显示,海岛棉GbF3’H基因沉默后其在海岛棉根、茎和叶中的表达量比空载体对照低,Gb F3’H、GbCHI和GbDFR这3种基因共沉默后其在海岛棉根、茎和叶中的表达量均比空载体对照低。病情指数调查结果显示,野生型<空载体对照相似文献   

Agronomic traits at the seedling stage,yield, and fiber quality in two cotton (Gossypium hirsutum L.) cultivars in response to phosphorus deficiency

ABSTRACT

Cotton is critical for phosphorus demands and very sensitive for its deficiency. However, identifying the effect of low-phosphorus tolerance on cotton growth, yield, and fiber quality by reducing phosphorus consumption. This may help to develop phosphorus-tolerant high-yielding cotton cultivars. In a two-year repeated (2015 and 2016) hydroponic experiment (using 0.01 and 1 mM KH₂PO₄), two cotton cultivars with phosphorus sensitivity (Lu 54; a low-phosphorus sensitive and Yuzaomian 9110; a low-phosphorus tolerant) were screened on the base of agronomic traits and physiological indices through correlation analysis, cluster analysis and principal component analysis from 16 cotton cultivars. Low phosphorus nutrition reduced the plant height, leaf number, leaf area, phosphorus accumulation and biomass in various organs of seedlings. The deficiency negatively affected the morphogenesis of seedlings, as well as yield and fiber quality. Further, these screened cultivars were tested in a pot experiment with 0, 50, 100, 150, 200 kg P₂O₅ ha^?1 during 2016 and 2017. It was found to have a significant (P< 0.05) difference in boll number, lint yield, fiber strength, and micronaire at the harvest. Furthermore, after collectively analyzed the characteristics of Lu 54 and Yuzaomian 9110, there were six key indices that could improve the low phosphorus tolerance of cotton cultivars. These were root phosphorus accumulation, stem phosphorus accumulation percentage, leaf and total biomass of seedlings, seed cotton weight per boll and fiber length.     

Isolation and Characterization of GhLEA3 Gene from Upland Cotton and Its Expression in Response to Low Temperature Stress

[Objective] Based on the analysis of cotton GhLEA3 gene structure and its expression model in chilling stress, we investigated the resistance function of GhLEA3 in response to cold stress. [Method] We cloned GhLEA3 from upland cotton by homologous sequence cloning way. We analyzed the properties of the GhLEA3, and constructed phylogenetic tree by bioinformatics analysis. We constructed the transient expression vector 35S∷GhLEA3-GFP by In-Fusion connection technology and studied the subcellular localization of GhLEA3. The expression of GhLEA3 gene in leaves of TM-1 at three-leaf stage under low temperature stress treatments (4 ℃, 24 h) was performed. Agrobacterium tumefaciens mediated floral dipping method was used to transform the gene with expression vector 35S∷GhLEA3-GFP into Arabidopsis thaliana wild type. Low temperature germination experiment at 4 ℃ was conducted to verify germination ability of the transgenic Arabidopsis thaliana T₃ seed. The transgenic Arabidopsis seedlings were treated at 4 ℃(low temperature), and all the leaves were taken to measure their electrical conductivity. [Result] The coding sequence of GhLEA3 gene is 1 218 bp, which encodes 405 amino acids. GhLEA3 protein includes 4 functional domains of PF02987. Phylogenetic analysis showed that the similarity of amino acid sequences between upland cotton GhLEA3 and Arabidopsis thaliana AtLEA3 was 52.6%. GhLEA3 might be mainly localized in vacuoles and small vesicles. GhLEA3 was up-regulated in leaves after low temperature treatment. The germination rate of transgenic Arabidopsis thaliana of T₃ generation at low temperature was significantly higher than that of wild type, and its electrical conductivity of leaves after low temperature treatment was significantly lower than that of wild type. [Conclusion] GhLEA3 belonged to the member of LEA3 family. Phylogenetic analysis showed that GhLEA3 had the closest relationship with AtLEA3. GhLEA3 was induced by low temperature stress and may play an important role in improving cold resistance.     

Identification of resistance to Aspergillus flavus infection in cotton germplasm

Natural resistance in cottonseed to Aspergillus flavus infection has not been explored to date. A green fluorescent protein (GFP) expressing A. flavus strain was used to assess the resistance of seed from 35 cotton varieties from Gossypium arboreum, G. barbadense, and G. hirsutum. Mature cotyledons devoid of seed coat were wounded, inoculated, and assessed for innate resistance to A. flavus infection. Of the initial 35 varieties tested, we observed a range of resistance to infection in representatives of all three species. A subset of 15 representatives was further analyzed. Within this group, G. arboreum cultivar A2 186 and G. hirsutum cultivar SA 1582 were most resistant to fungal infection. The most susceptible cultivar in this group was G. hirsutum SA 1595. The remaining 12 representatives tested in the secondary screen (3 G. arboreum, 3 G. barbadense, and 6 G. hirsutum lines) exhibited intermediate resistance. We did not observe any relationship between species and resistance. Within each species, there was a range of responses to fungal infection. Future studies using this methodology to screen additional diploid and tetraploid cotton lines may enable us to identify naturally resistant germplasm that can be used to develop cotton with enhanced resistance to fungal infection.     

Electron Transport Through Photosystem II Is Not Limited By A Wide Range of Water Deficit Conditions In Field‐Grown Gossypium hirsutum

Despite exhaustive literature describing drought stress effects on photosynthesis in Gossypium hirsutum, the sensitivity of photosynthetic electron flow to water deficit is heavily debated. To address this, G. hirsutum plants were grown at a field site near Camilla, GA under contrasting irrigation regimes, and pre‐dawn water potential (Ψ_PD), stomatal conductance (g_s), net photosynthesis (P_N), actual quantum yield of photosystem II (Φ_PSII) and electron transport rate (ETR) were measured at multiple times during the 2012 growing season. Ψ_PD values ranged from ?0.3 to ?1.1 MPa. Stomatal conductance exhibited a strong (r² = 0.697), sigmoidal response to Ψ_PD, where g_s was ≤0.1 mol m^?2 s^?1 at Ψ_PD values ≤ ?0.86 MPa. Neither Φ_PSII (r² = 0.015) nor ETR (r² = 0.010) was affected by Ψ_PD, despite exceptionally low Ψ_PD values (?1.1 MPa) causing a 71.7 % decline in P_N relative to values predicted for well‐watered G. hirsutum leaves at Ψ_PD = ?0.3 MPa. Further, P_N was strongly influenced by g_s, whereas ETR and Φ_PSII were not. We conclude that photosynthetic electron flow through photosystem II is insensitive to water deficit in field‐grown G. hirsutum.