棕色棉茎尖基因枪遗传转化

为了建立高效的棉花茎尖遗传转化体系,以棕色棉品种新彩棉5号为材料,采用茎尖培养和基因枪遗传转化方法,通过对凝固剂种类、活性炭含量、卡那霉素浓度、渗透处理时间、轰击压力、轰击距离等因素的优化,建立了棕色棉新彩棉5号茎尖基因枪遗传转化技术体系。结果表明,不同凝固剂种类、活性炭含量、卡那霉素浓度、渗透处理时间、轰击压力和轰击距离对棕色棉茎尖转化率有明显影响。最佳的遗传转化体系为:前渗处理4 h,轰击压力7.586 MPa(1100 psi),轰击距离9 cm,轰击后恢复培养4 d,然后在MSB+8 g·L~(-1)琼脂+125mg·L~(-1)卡那霉素筛选培养基中筛选25 d。获得抗性芽后,在添加1 g·L~(-1)活性炭的生根培养基中诱导抗性芽生根,并获得抗性再生植株。通过PCR和q PCR检测,5200个茎尖共获得16株阳性转基因植株,转化率为0.31%。该技术体系的建立为棕色棉的遗传转化奠定了基础,为彩色棉育种提供了新的材料。     

A Regeneration System for Cotton Variety Xinluzao 45 via Somatic Embryogenesis

[Objective] The purpose of this study was to expand upland cotton genotypes suitable for regeneration via somatic embryogenesis, so as to provide excellent receptor materials for the genetic transformation of cotton. [Method] Six different hormone combinations were studied to establish a somatic embryo regeneration system for hypocotyl explants of Xinluzao 45, a widely planted variety in Xinjiang Province. [Result] Faint yellow callus was induced on a medium consisting of hormone combinations C1 (0.1 mg·L^－1 acetic acid dichlorobenzene oxide [2, 4-D] + 0.1 mg·L^－1 kinetin [KT]) or C5 (0.1 mg·L^－1 2, 4-D + 0.5 mg·L^－1 indo-3-butyric acid + 0.1 mg·L^－1 KT). The generated callus was able to successfully induce embryogenic callus on hormone-free medium containing doubled KNO3. The embryogenic callus then developed into somatic embryos and regenerated plants. Maximum rates of callus induction and regeneration were 92.2% and 40.3%, respectively. Although no significant difference was observed in the rate of callus induction of Xinluzao 45 hypocotyls between hormone combinations C1 and C5, the incidence of induced embryonic callus was higher on C1 than on C5. [Conclusion] The somatic embryo regeneration system developed in this study provides technical support that will facilitate Agrobacterium-mediated transformation of upland cotton Xinluzao 45.     

铜基营养保护剂对棉花生长和产量的影响

2013―2014年采用盆栽试验研究了不同种类及浓度的铜基营养保护剂对棉花生长、产量及防病效果等的影响。结果表明:4种铜基营养保护剂在喷施质量浓度为1 g·L-1和2 g·L-1时,与喷施清水相比,皮棉产量分别增加4.22%~13.38%和21.89%~38.36%;增加了棉株地上部干物质质量,最高达23.96%;降低了棉花病情指数26.75%~40.91%和22.78%~37.78%,但处理间差异不显著。因此,在棉花上喷施铜基营养保护剂既可促进棉花生长,增加棉花产量,又可提高棉花的防病效果。     

Effects of Glyphosate on Soil Salinization and Alkalization in Cotton Fields

We analyzed the effects of glyphosate to the index of salinization and alkalization by simulation in soils collected from cotton fields in Anyang, Henan and Anqing, Anhui, China, in 2014. The results showed that application of glyphosate changed electrical conductivity (EC), total alkalinity (TAL), exchangeable sodium percentage (ESP), sodium adsorption ratio (SAR) and pH of soil in both regions，compared with the blank control. With increased frequency of glyphosate application and increased concentration applied, all indices increased in the Anqing cotton fields soil which were treated with concentrations of 2.5-20 g·L^－1 . However, EC, TAL and ESP increased dynamically in Anyang soil to which high concentrations of 20 g·L^－1 were applied only four consecutive times. After different glyphosate treatments, the soil pH value showed an upward trend but smaller changes. Therefore, after repeated use of glyphosate to soils of two different region soils, although the changes of all indices were within the range of non-saline and non-sodic soils, the soils affected by salt in the glyphosate aqueous solution tended towards saline-sodic soils.     

脱落酸对棉花体细胞胚胎发生的影响

【目的】研究脱落酸(Abscisic acid, ABA)对棉花体细胞胚胎发生过程中下胚轴脱分化和再分化的影响,优化体细胞胚胎发生体系和初步解析脱落酸调控棉花体细胞胚胎发生分子机制。【方法】以棉花品种中棉所24(CCRI 24)下胚轴为外植体,设置5个ABA浓度0、0.02、0.04、0.06、0.08μmol·L^-1,分别以A0、A1、A2、A3、A4表示,添加至MSB(MS培养基+B5维生素)培养基诱导愈伤和胚性愈伤,研究ABA对棉花下胚轴初始细胞脱分化、愈伤组织诱导和胚性愈伤组织诱导的影响。【结果】ABA促进下胚轴初始细胞脱分化;显著提高愈伤组织的脱分化率和增殖率;0.02μmol·L^-1ABA显著提高胚性愈伤分化率,0.04~0.08μmol·L^-1ABA显著降低胚性愈伤分化率。ABA处理后胚性愈伤和非胚性愈伤的增殖率均显著提高且质地受到影响。0.02~0.08μmol ABA处理下,LBD和LBD在愈伤起始期上调表达。0.02μmol·L^-1ABA处理下,在愈伤增殖早期和中期BBM、LEC1和AGL15上调表达,愈伤增殖后期FUS3、LEA、ABI3基因上调表达。【结论】脱落酸调控的棉花体细胞胚胎发生与相关标记基因的时空性表达密切相关,这些基因表达水平的增加是ABA调控愈伤和胚性愈伤分化的分子基础。     

Effects of Exogenous 24-epibrassinolide on Photosynthetic Physiology of CottonSeedlings under Low Temperature

[Objective] The objective of this study was to investigate the effects of exogenous 24-epibrassinolide (EBR) on the photosynthetic physiology of cotton seedlings under low temperature and to provide basis for improving the cold tolerance of cotton by using EBR as growth regulator. [Method] Taking CCRI 60, Lumianyan 28 and Simian 3 as materials, a field experiment was carried out in Institute of Cotton Research of CAAS(Anyang county, Henan province). Before the first low temperature treatment, the cotton seedlings were sprayed with distilled water (Control) and different concentrations of EBR (0.1 mg·L^－1 and 0.2 mg·L^－1), respectively. After 3 days, the relative electrical conductivity, chlorophyll content, rapid chlorophyll fluorescence induction kinetic curve (OJIP) and fluorescence parameters were measured. [Result] Under low temperature, the relative conductivity of CCRI 60, Lumianyan 28 and Simian 3 spraying with EBR decreased by 17.7%～32.8% compared with control, and there was no significant difference between CCRI 60 and Lumianyan 28 in different concentrations of EBR treatments, butthe relative conductivity of Simian 3 treating with 0.2 mg·L^－1 EBR was significantly lower than those treatments with 0.1 mg·L^－1 EBR . The chl a and chl b contents increased by 9.7%～32.6% and 15.0%～18.9%, respectively. The maximum photochemical efficiency of photosystemⅡ (F_v/F_M) and photosynthetic performance index on absorption basis(PI_ABS) increased significantly. PI_ABS of CCRI 60 increase the maximum by 75.6% using 0.1 mg·L^－1 EBR. Lumianyan 28 and Simian 3 increased the maximum by 101.1% and 265.6% using 0.2 mg·L^－1 EBR, respectively; Absorbed photon flux per cross section (ABS/CSm), electron transport flux (further than QA) per active reactive center (ETo/RC) and probability for electron transport (φEo) are significantly increased. [Conclusion] Exogenous EBR can enhance the ability of low temperature tolerance of cotton seedlings and alleviate the inhibition of photosynthesis in cotton at low temperature. The study showed that 0.1 mg·L^－1 EBR performs well in CCRI 60 and 0.2 mg·L^－1 in Lumianyan 28 and Simian 3.     

Effects of Melatonin on the Antioxidant Enzyme Activities and Seed Germination of Cotton (Gossypium hirsutum L.) under Salt-Stress Conditions

[Objective] Cotton (Gossypium hirsutum L.) seed germination, in which melatonin plays an important regulatory role, is seriously affected by soil salinization. Cotton seed germination, antioxidant enzyme activity levels and other physiological indicators were analyzed to clarify the regulatory effects of exogenous melatonin on cotton seed germination under salt-stress conditions. [Method] The experiment was conducted in a greenhouse of Hebei Agricultural University in Baoding City, Hebei Province from 2018 to 2019 using Guoxin Cotton 9 as the material. The germination rate, germination potential, seed biomass after germination, superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT) activities, as well as the malondialdehyde (MDA) content, of cotton seeds were measured after exposure to different melatonin concentrations (0, 10, 20, 50 and 100 μmol·L^－1) and 150 mmol·L^－1 NaCl treatments. [Result] After germination, the germination rate, germination potential, radical length and seed biomass significantly decreased, as did the SOD, POD and CAT activities. However, the MDA content significantly increased under salt-stress conditions. Low exogenous melatonin concentrations (10 and 20 μmol·L^－1) increased the germination potential, germination rate and biomass of cotton seeds, and promoted the elongation of radicals; however high melatonin concentrations (50 and 100 μmol·L^－1) inhibited cotton seed germination and decreased seed biomass after germination. Low melatonin concentrations (10 and 20 μmol·L^－1) increased the SOD, POD and CAT activities and decreased the MDA content in cotton seeds. [Conclusion] Low melatonin concentrations could promote germination and improve the salt tolerance of cotton seeds, while high melatonin concentrations could inhibit their germination. A 20 μmol·L^－1 melatonin concentration is appropriate for regulating cotton seed germination under salt-stress conditions.     

大草蛉化学感受蛋白基因CpalCSP3组织表达谱及气味结合特性分析

昆虫化学感受蛋白(Chemosensory protein,CSP)具有结合和运载气味分子穿过感器淋巴液到达嗅觉受体的作用,但尚未见有关大草蛉(Chrysopa pallens)CSP功能的报道。本研究克隆得到大草蛉Cpal CSP3基因,利用RT-q PCR技术测定了其在成虫各组织的表达谱,发现Cpal CSP3基因主要在雌雄虫的触角和头部表达,暗示该基因参与了大草蛉的嗅觉及味觉感受。为探讨其嗅觉功能,利用原核表达系统和荧光竞争结合试验测定了Cpal CSP3蛋白与84种气味物质的结合能力。结果表明,Cpal CSP3蛋白与橙花叔醇、(+)￣雪松醇和β￣紫罗兰酮具有很高的结合能力,结合常数(Ki)为8.79~9.79μmol·L-1;与根皮苷、月桂醛、乙酸香叶酯和α￣蒎烯也有较高的结合能力,Ki为11.54~19.06μmol·L-1。分析认为,Cpal CSP3蛋白可能在大草蛉对这些气味的嗅觉感受中起作用,使得大草蛉能对害虫寄主植物有效定向进而搜寻到猎物。     

Applying Plant Growth Regulators at Early Stages Affects Maturation of Cotton

[Objective] Early initiation and early maturity are the foundation of high yield and good quality of cotton. The purpose of this study is to determine the effects of plant growth regulators applied at the seedling and squaring stage on the early initiation of flower bud and the rate of the opened cotton boll (ROCB) during later development period, and to provide practical measures for hastening the maturity of cotton. [Method] Several plant growth regulators were applied from cotyledonary to squaring stage under greenhouse and field conditions, water was used as the control. The first fruiting branch node (indicating the initiation of flower bud), the number of bud prior to blooming and the ROCB at mid-term of boll maturation period (23 September, 2017) were compared among treatments. [Result] Under greenhouse conditions, gibberellic acid (GA₃) applied at the cotyledonary stage with 140 μmol·L^－1 as well as the three consecutive applications of sodium nitrophenolate (CSN, 2.23 μmol·L^－1) at the cotyledonary, two-leaf and four-leaf stage made the first fruiting branch node move down by about 0.9 nodes. In field experiments, the application of gibberellin₄₊₇(GA₄₊₇, 288 and 576 μmol·L^－1) at the cotyledonary stage significantly decreased the first fruiting branch node by about 0.4 nodes. Also, the application of 6-benzylaminopurine (6-BA, 44.4 μmol·L^－1) at the three-leaf stage significantly decreased the first fruiting branch node by 0.2 nodes. However, there was no significant correlation between the first fruiting branch node and the ROCB in late September. Moreover, the application of Brassinolide (BR, 0.10 μmol·L^－1) at the bud stage increased the ROCB in late September, which was mainly associated with the increased boll set in the lower and middle fruiting branches. [Conclusion] The reasonable distribution of bolls (concentrated in the lower and middle fruiting branches as well as inner fruiting sites) is more important for the earliness of cotton than lowering the first fruiting branch node.     

Effects of the Seed Film Coating Salicylic Acid on the Chilling Tolerance of Cotton Seedlings

The objective of this study was to investigate the effect of the different concentrations of the seed film coating salicylic acid on the cold tolerance of cotton seedlings using the sand culture method. The physiological response of cell membrane permeability, protective enzyme activity and osmotic adjustment of cotton seedling were measured before and after at low temperature stress and at normal temperature. The results showed that 1-10 mmol·L^－1 of seed film coating with salicylic acid seed coating treatment could effectively alleviate damage to cotton seedlings at low temperature, and the accumulation of the relative electrical conductivity and malondialdehyde were reduced on cotton seedlings. The activity of superoxide dismutase, peroxidase, catalase, and soluble protein, soluble sugar and proline contents increased sharply. These results indicate that film coating with salicylic acid can alleviate the effects of low temperature stress on seedling growth inhibition and improve the cotton seedlings by providing chilling tolerance, and the optimum treatment concentration salicylic acid is 5 mmol·L^－1.     

不同咸水利用方式对棉花叶绿素荧光参数及土壤盐分的影响

【目的】本研究旨在科学合理地利用浅层咸水资源。【方法】依托短期定位试验开展了在第3年和第4年不同咸水利用方式下(CK:造墒和蕾期灌淡水;T1:造墒和蕾期灌咸淡混配矿化度3 g·L^-1的微咸水;T2:淡水造墒蕾期灌矿化度5 g·L^-1咸水;T3:造墒和蕾期灌矿化度5 g·L^-1咸水;T4:淡水造墒蕾期不灌水)棉花长势、叶绿素荧光参数、土壤盐分累积及其运移的变化。【结果】T1和T2处理的齐苗率、株高、干物质质量、叶面积指数、叶绿素荧光参数、产量及霜前花率与CK无显著差异,土壤盐分含量较CK有所增加,但未对棉花生长产生明显抑制。T3处理的棉花长势指标、叶绿素荧光参数较CK显著降低,0~100 cm土壤盐分含量较CK明显增加。【结论】从土壤质量安全和咸水高效利用的角度考虑,连续4年用3 g·L^-1的咸淡混合水灌溉或用淡水与5 g·L^-1的咸水轮灌不仅能节约淡水,且不影响棉花产量。本研究结果为当地在棉花生产中安全利用咸水提供技术参考。     

改进的TPS法:一种棉花叶片DNA快速提取方法

分子标记辅助选择需要大批量的DNA样品,快速提取DNA的方法是提高选择效率的关键。研究比较多种植物DNA的提取方法,通过优化改进,建立了适于棉花叶片基因组微量快速提取的新方法。该方法增加缓冲液预处理及在TPS缓冲液中添加PVP40,采用PCR板批量取样、微型手电钻磨样,省去了蛋白抽提、DNA沉淀、洗涤、风干、溶解等步骤,免去氯仿和异戊醇的使用。本方法具有低成本、快速、高通量、操作简单等特点,而且能确保提取的棉花DNA的浓度和纯度达到要求,满足大批量样本SSR分子标记辅助育种工作的需要。     

种植密度和缩节胺调控对麦后直播棉产量和冠层特征的影响

2013―2014年以早熟棉(中棉所50)为材料,采用裂区设计,在江苏省南京市研究了种植密度(7.50万、9.75万和12.00万株·hm~(-2))和缩节胺(DPC)调控(0,52.5和105.0 g·hm~(-2))对麦后直播棉产量和冠层特征的影响。结果表明:皮棉产量在不同种植密度下以12.00万株·hm~(-2)处理最低,在不同DPC用量水平下以0 g·hm~(-2)处理最低;种植密度与DPC调控存在互作效应,以种植密度9.75万株·hm~(-2)、DPC用量52.5~105 g·hm~(-2)处理产量较高,且产量构成中以铃数对产量的直接效应最大。对冠层特征影响表明,下部果枝夹角和长度随种植密度增加而降低,而中、上部果枝的夹角和长度、叶面积指数均以种植密度9.75万株·hm~(-2)处理较高;不同部位果枝夹角和长度、叶面积指数均随DPC用量增加而降低,而透光率则相反。相关分析表明,下部果枝夹角大、中部果枝较长及上部果枝夹角小且叶面积指数和透光率较高,有利于提高产量和霜前花率。综上,该棉区麦后直播棉种植密度9.75万株·hm~(-2)、DPC用量52.5~105 g·hm~(-2)(蕾期、开花期和打顶后用量比例为1∶2∶4),有利于改善棉花冠层特征,实现早熟高产。     

外源调节物质对棉花幼苗耐寒生理特性的效应

以棉花幼苗为试材,叶面分别喷施外源调节物质水杨酸(SA)、壳聚糖(CTS)、水杨酸+壳聚糖(SCM)、水杨酸+壳聚糖+硝酸钙(SCCa M)后,在5℃低温下进行处理,分别于0、24 h、48 h及恢复处理24 h时测定相对电导率、MDA含量、叶绿素含量,SOD、POD、CAT的活性和可溶性蛋白、可溶性糖、脯氨酸的含量。结果表明:低温胁迫前用0.7 mmol·L-1水杨酸、0.6 mmol·L-1壳聚糖及复配制剂处理棉花幼苗,其叶片的SOD、POD、CAT活性和叶绿素、可溶性蛋白、可溶性糖、脯氨酸含量与对照相比均有不同程度的提高,相对电导率和MDA含量有所降低;复配制剂处理效果优于水杨酸和壳聚糖单独施用的效果,而水杨酸、壳聚糖、硝酸钙三元复配的效果比水杨酸和壳聚糖两元复配的效果好。表明水杨酸、壳聚糖和硝酸钙在诱导棉花幼苗耐寒性中具有协同效应。     

Effects of Exogenous Proline on the Growth,Physiological Characteristics,and Proline Metabolism of Cotton Seedlings under Boron Deficiency Stress

[Objective] This study investigates the effects of exogenous proline on the growth, physiological responses, and proline metabolism of cotton (Ekang No.8) seedlings treated with varying boron (B) concentrations. [Method] We conducted a randomized experiment with six treatments using the hydroponic method in a greenhouse at Huazhong Agricultural University. We applied three different concentrations of boron, including 0.02 μmol·L^－1 (low-concentration B), 2.5 μmol·L^－1 (medium-concentration B), and 100 μmol·L^－1 (sufficient-concentration B). The first two are boron-deficient treatments. We also applied an exogenous proline treatment at 20 μmol·L^－1(0 μmol·L^－1 as control). When any significant difference among the treatments were observed, the related indicator was measured. [Result] The results showed that exogenous proline inhibited the growth of cotton seedlings under sufficient-concentration B treatment while promoting the absorption of B by roots under low-concentration B treatment. The application of exogenous proline to the seedlings under low-concentration B treatment reduced the contents of proline and H₂O₂ in leaves but increased the accumulation of MDA and H₂O₂ in roots. The activities of SOD and antioxidant enzymes (APX) in the roots and leaves were dramatically enhanced. Conversely, POD activity reduced significantly and there was no significant change in CAT activity relative to low-concentration B treatment. More importantly, we found that the application of exogenous proline under B deficiency increased the activities of P5CS, P5CR, OAT (synthetase), and PRODH (degrading enzyme) in proline metabolic pathways. [Conclusion] Applying exogenous proline under sufficient-B concentration inhibits growth. The application of proline to seedlings under low-B concentration promotes the absorption of B by roots, increases the activity of APX, and decreases the membrane lipid peroxidation in leaves. A B deficiency leads to proline accumulation in plants. The addition of proline under low-B concentration can reduce the proline content in leaves, which is caused by affecting the critical synthetase and catabolic enzyme activities in the proline metabolism pathway (Glu and Orn pathway). The main reason for this occurrence is the significant increase of proline dehydrogenase (PRODH) activity.     

In vitro Antifungal Activity and Antifungal Mechanism of Essential Oil from Dill (Anethum graveolens L.) on Verticillium dahliae Kleb.

The objectives of the present study were to assess the antifungal effect of dill seed essential oil (EO) against V. dahliae and describe its mechanism of action. The microbroth dilution method was used at the minimal inhibitory concentration (MIC) and the dry mycelium weight and spore germination of the tested V. dahliae were also determined in a liquid culture to evaluate the anti-V. dahliae activity in vitro. The results showed EO was active against V. dahliae and MIC was 0.625 μL·mL^－1. Mycelial growth, mycelium weight and spore germination were inhibited by the EO in a dose-dependent manner. The results show that upon treatment of cells with EO, propidium iodide penetrated V. dahliae through a lesion in its plasma membrane. Thus, our results demonstrated that EO could be a potential source of plant-derived fungicides to control V. dahliae.     

Interaction of Plant Density with Mepiquat Chloride Affects Plant Architecture and Yield in Cotton

[Objective] The effect of planting density and mepiquat chloride (DPC) on cotton plant architecture, growth, yield, and quality at Anyang City, Henan Province, China, was studied. [Method] Field experiments with cotton variety Lumianyan 28 were conducted with five planting densities (15 000, 45 000, 75 000, 105 000, and 135 000 plants·hm^－2) and application of DPC at three concentrations (0, 195, and 390 g·hm^－2). [Result] Increasing cotton plant density resulted in increased internode length and plant height but also caused the decrease of inclination of fruiting branches and leaves as well as elevated dry matter allocation to leaves and fruiting branches, which led to a decrease in dry matter accumulation. Application of DPC reduced the azimuth angle of fruiting branches and plant height, but increased the insertion angle of fruiting branches with the main stem, leaf length, and petiole length. Planting density and DPC treatment showed a significant interaction on fruiting branch insertion angle, plant height, stem diameter, and dry matter allocation to fruits and leaves. The interaction of DPC and planting density had a complementary effect on the spatial distribution of cotton-yielding bolls. The final dry matter was highest (14 362 kg·hm^－2) at the planting density of 105 000 plant·hm^－2 and DPC application of 390 g·hm^－2, which resulted in the highest seed yield (3 257 kg·hm^－2). [Conclusion] For maximization of cotton yield and quality, a plant density of 75 000 to 105 000 plants·hm^－2 and DPC application of 195 to 390 g·hm^－2 in the Yellow River cotton-producing region is recommended. The results may help to optimize labor-saving cotton management and to generate a plant architecture suitable for mechanical harvesting in the Yellow River cotton-producing region.     

防治烟蓟马的生物源农药筛选及其对解毒酶活性影响

【目的】筛选防治烟蓟马(Thrips tabaci Lindeman)的高效生物源农药,探究其对烟蓟马3种主要解毒酶细胞色素P450单加氧酶(Cytochrome P450 monooxygenase,P450)、谷胱甘肽S-转移酶(Glutathione S-transferase,GST)和羧酸酯酶(Carboxylesterase,CarE)活性的影响。【方法】以棉田采集并室内稳定饲养的烟蓟马为供试虫源,用浸叶法测定7种生物源农药对烟蓟马若虫的毒力,选择活性好的3种药剂开展田间药效试验,并在这3种生物源农药亚致死浓度(LC25)剂量处理后测定烟蓟马体内3种主要解毒酶活性。【结果】(1)7种生物源农药对烟蓟马的毒力从高到低依次为乙基多杀菌素、多杀霉素、苦参碱、印楝素、除虫菊素、鱼藤酮、藜芦碱。乙基多杀菌素、多杀霉素和苦参碱对烟蓟马的致死中浓度(LC50)分别为0.011 mg·L^-1、0.274 mg·L^-1、1.479 mg·L^-1。(2)田间药效试验发现,每666.7 m2用2 g、4 g、6 g的60 g·L^-1乙基多杀菌素悬浮剂和10 g的5%(质量分数,下同)多杀霉素悬浮剂处理,施药后7 d防治效果均在90%以上,优于1.3%苦参碱水剂和对照药剂30%噻虫嗪悬浮剂各剂量的防效。(3)LC25剂量处理烟蓟马48 h后,乙基多杀菌素处理的烟蓟马体内P450、GST和CarE活性升高,苦参碱和多杀霉素处理后CarE活性升高,噻虫嗪处理后P450和CarE活性升高。【结论】乙基多杀菌素、多杀霉素和苦参碱对烟蓟马的毒力较高,可用于防控棉田烟蓟马;CarE可能参与了烟蓟马对多杀霉素和苦参碱的解毒代谢,3种主要解毒酶可能均参与了烟蓟马对乙基多杀菌素的解毒代谢。     

Development of a Biological Complex Seed-Coating Agent to Control Verticillium Wilt

[Objective] The aim of this study is to make use of the antagonism and growth promoting ability of Bacillus methylophicus strain LH-L3, and add it into the chemical seed-coating agent as an active ingredient. [Method] The optimum formula (agent No.4: 2.5% fludioxonil＋10% thiram＋10% carbofuran＋109 cfu·mL^－1 LH-L3) was obtained based on the sterilization effect of four seed coating agents on infected seed. Then the survival ability of the strain in agent No.4 and its shelf life were identified by plate test. Finally, the disease and pest control effects were tested in the greenhouse pot experiment. [Result] The results showed that the agent No. 4 had the best control effect, and 1.1×105 cfu·mL^－1 of LH-L3 after storage at room temperature for 6 months. Compared with the control, the formula No. 4 increased 11.54% emergence rate, 18.74% plant type height and 20.91% leaf width, respectively, in the 15th day after emergence, and decreased 65.42% and 48.56% of Verticillium wilt at day 2 and day 5 after the disease occurred. The cotton seeds coated with formula No. 4 reduced 83.07% of plant and 92.33% of leaf damaged by Sylepta derogata, compared with the negative control without seed coating. [Conclusion] Strain LH-L3 and chemical pesticide compound as a seed coat agent is completely feasible. And the developed biological complex seed-coating agent has better control effect on Verticillium wilt and Sylepta derogate.     

Generation and Glyphosate Resistance Analysis of G10eve Transgenic Cotton

[Objective] Novel germplasm of cotton was generated for development of cultivars with high resistance to glyphosate. [Method] The glyphosate resistance gene G10eve was preliminarily analyzed using bioinformatics methods and introduced into 'Coker 312' via Agrobacterium-mediated transformation. The expression of G10eve was estimated by PCR (Polymerase chain reaction) and qRT-PCR (Quantitative real-time PCR). The content of shikimic acid and tolerance to glyphosate were determined in response to application of glyphosate to 'Coker 312' and transgenic plants. [Result] A total of 28 independent positive transgenic lines were obtained. The transformation efficiency and the rate of seedling differentiation were up to 49.3% and 40.6%, respectively. PCR and qRT-PCR analysis showed that G10eve was integrated into the cotton genome, and the G10eve expression level differed significantly among the transgenic lines. At the cotyledon stage, the transgenic cotton showed resistance to 8 mL·L^－1 glyphosate, whereas 'Coker 312' showed phytotoxicity to the 2 mL·L^－1 glyphosate treatment. No significant accumulation of shikimic acid was detected in transgenic plants before or after glyphosate treatment, which indicated the typical physical characteristic of glyphosate resistance. [Conclusion] Overexpression of G10eve in cotton improved resistance to glyphosate. Thus, novel germplasm of cotton resistant to glyphosate was generated.