基于光谱红边参数的棉花黄萎病叶片氮素含量诊断研究

 以黄萎病胁迫下棉花叶片为供试材料,分析了黄萎病棉叶氮素含量(LNC)与光谱红边参数间的关系,建立了黄萎病棉叶LNC(Leaf nitrogen content) 的光谱红边参数诊断模型。结果表明：(1) 随着黄萎病严重程度的增加,棉叶LNC逐渐减小,且差异显著;（2）黄萎病棉叶红边参数红边位置(REP) 、红边振幅（Dr）、红谷位置(Lo) 、红边深度（Depth₆₇₂）和红边面积(Area₆₇₂) 均减小,红边宽度(Lwidth) 增加,且Area₆₇₂的值减小的幅度最大,Dr减小的幅度最小,Lwidth的值增加的幅度较大; (3) 黄萎病棉叶LNC含量均与红边参数REP、Lo、Depth₆₇₂和Area₆₇₂呈极显著正相关,与Lwidth呈极显著负相关,与Dr未达显著相关;(4)基于红边参数建立的黄萎病棉叶LNC含量的诊断模型均达到极显著水平（P<0.01）,其中以Area₆₇₂为自变量建立的黄萎病棉叶LNC的诊断模型的精度最高,R²超过0.7,RMSE小于0.6,RE小于0.007,能很好地诊断黄萎病棉叶LNC。     

文山辣椒成熟期不同颜色果实的色素含量

为揭示云南文山州辣椒成熟期果色多样性和独特性的化学基础,用分光光度法研究了该州辣椒成熟期紫色、红色、金色和杂色果实的叶绿素(chlorophylls,Chls)、类胡萝卜素(Car)和总花色苷含量。结果表明:文山辣椒成熟期不同颜色果实的叶绿素a(Chl a)和总叶绿素[Chl(a+b)]含量和叶绿素a/b(Chl a/b)及总叶绿素/类胡萝卜素[Chl(a+b)/Car]特征均为紫色杂色红色金色,叶绿素b(Chl b)含量特征为紫色杂色金色红色,Car含量特征为金色红色杂色紫色,总花色苷含量(TAC)特征为紫色红色金色杂色,此外,4种颜色果实Chl a、Chl b和Chl(a+b)含量间的差异均达到极显著水平,但Car含量和TAC间的差异则均未达到显著水平。因此,文山辣椒成熟期果实颜色差异主要决定于Chl含量,叶绿素和花色苷共同贡献于果实的紫色,Car含量则决定了果实的金色和红色。本研究可为文山州辣椒果色呈现机理的研究和新奇果色品种的选育提供参考。     

利用叶片高光谱指数预测水稻群体叶层全氮含量

通过测定叶片高光谱来快速估测整个水稻叶层全氮含量对于水稻氮素诊断有重要意义。本文通过连续3年不同施氮水平和不同品种类型的4个大田试验,分生育期同步测定了不同叶位叶片的高光谱反射率及叶层全氮含量,并系统分析了叶片水平多种高光谱指数与水稻叶层全氮含量的定量关系。结果表明,不同叶位叶片的光谱反射率与叶层全氮含量的相关程度不同,顶二叶(L2)表现最好、顶三叶(L3)次之,而L2和L3的平均光谱(L23)有助于进一步提高光谱指数的敏感性,是估测叶层氮含量的适宜叶位组合。绿光560nm和红边705nm波段附近光谱反射率与叶层全氮含量呈极显著负相关关系,两者分别与近红外波段组合而成的光谱比值指数可较好地监测水稻叶层全氮含量,其中绿光、红边窄波段比值指数SR(R780,R580)和SR(R780,R704)表现较好,与叶层全氮含量的决定系数分别为0.887和0.884;独立试验数据检验的RMSE分别为0.216和0.235。将上述2个窄波段比值指数中的近红外、绿光波段和红边波段宽度分别扩展至100、20和10nm,从而构建的宽波段比值指数SR[AR(750-850),AR(568-588)]和SR[AR(750-850),AR(699-709)]与叶层全氮含量相关性仍具有较高水平,线性回归模型的拟合精度(R2)为0.886和0.883,检验RMSE值分别为0.218和0.237。从而在叶片水平,确立了适于叶层全氮含量估测的基于绿光、红边与近红外波段的比值组合和波段适宜宽度。     

棉花功能叶片色素含量与高光谱参数的相关性研究

叶片色素状况是评价植株光合能力、监测生长状况和预测产量潜力的重要指标,高光谱遥感技术为快速无损监测作物叶片色素提供了有效手段.本研究以4个棉花品种在3个施氮水平下的2年田间试验为基础,通过测定棉花(Gossypium hirsutum)功能叶片的高光谱反射率及对应的色素(叶绿素a、叶绿素b、叶绿素a b、类胡萝卜素)含量,定量分析了叶片高光谱参数与色素含量之间的相关关系.结果表明,与棉花功能叶片各色素指标相关性比较好的高光谱波段主要分布在500～700 nm;由敏感波段构建的光谱指数与各色素指标的相关性均在0.50以上;且红边最小值(Lo)可以作为共同的高光谱指数来估测不同棉花品种不同氮素水平下功能叶片的叶绿素总量(组合品种的R2为0.67).因此,通过高光谱参数来估算棉花功能叶片色素含量是可行的.     

玉米叶片光合生理与抗旱性关系研究

在水分胁迫条件下,对玉米叶片净光舍速率、叶绿素含量及叶绿素荧光参数的变化进行了研究.结果表明:玉米叶片净光合速率(Pn)、叶绿素a(Chl a)、叶绿素b(Chl b)、总叶绿素含量(Chl)、叶绿素a/b比值(Chl a/b)、最大荧光(Fm)和PsⅡ光化学效率(Fv/Fm)比对照均有所下降,基础荧光(Fo)增加;Pn和Ctd a/b比值占对照百分率与品种抗旱性呈极显著正相关(r=0.8807**,P<0.01;r=0.8804**,P<0.01),Fo占对照百分率与品种抗旱性则呈显著负相关(r=-0.764 0*,P<0.05),Fm和Fv/Fm占对照百分率与品种抗旱性则呈显著正相关(r=0.711 6*,P<0.05;r=0.772 3*,P<0.05).因此,净光合速率、叶绿素a/b比值、Fo、Fin和Fv/,m可以用来评定不同玉米品种的抗旱性.     

棉花高光谱及其红边特征(Ⅱ)

通过大田和室内试验,测定了2个棉花品种的冠层、完全展开倒1、3叶在不同时期的高光谱反射率及对应叶片的叶绿素、类胡萝卜素含量。结果表明:棉花冠层光谱红边具有“双峰”和“红边平台”现象,且红边位置λ_(red)位于695～720nm之间,红边幅值Dλ_(red)和红边面积S_(red)有“红移”和“蓝移”现象;叶面积指数、鲜叶重和干叶重与冠层光谱红边参数λ_(red)、Dλ(red)、S_(red)之间存在显著相关,叶片叶绿素和类胡萝卜素含量与其反射光谱的λ_(red)、Dλ(red)、S_(red)也有显著相关。     

基于吸收、透射和反射光谱预测水稻叶绿素含量研究

选择基于吸收率和透射率的叶绿素含量定量反演波段组合,构建叶绿素含量光谱估测模型寻找基于吸收、透射和反射光谱预测叶绿素含量的波段。以3个水稻品种临稻11,圣稻13和阳光200为材料,进行田间实验。比较水稻叶片吸收、反射及透射光谱曲线和一阶导数光谱曲线,发现440、480、630nm和681nm为叶绿素吸收峰的实际发生波段位置,其中630nm波段处的叶片光谱吸收率(A)、透射率(T)和反射率(R)之间相关性最好。比较三者之间的相关性,吸收率与透射率的相关性最强。630nm波段处的叶片光谱吸收率、透射率和反射率与叶绿素含量之间的相关性均达到极显著水平。回归分析表明基于440、480nm和681nm3个波段光谱吸收率线性模型,440、480nm和630nm3个波段光谱透射率线性模型估测叶绿素a含量,480、630nm和681nm3个波段光谱透射率线性模型估测叶绿素b含量,与单独使用630nm光谱变量估测叶绿素含量比较,在4个生育期估测精度均有显著提高,其中以叶绿素a和叶绿素总量的估测效果最好。     

基于棉花冠层光谱的土壤氮素监测研究

通过连续2年小区氮肥试验,在棉花不同生育期采集冠层高光谱数据并同步测定土壤氮含量,分析棉花冠层高光谱参数与土壤氮含量间的关系,建立基于植株冠层光谱的土壤氮含量估算模型。结果表明:土壤全氮含量随着施氮水平的增加而增加,且差异显著;基于棉花不同时期冠层光谱构建的14种光谱参量与土壤氮含量间的相关性有显著差异。其中,利用冠层光谱参数P_Area1100、Depth980、Area672、PPR(550,540)建立的土壤氮含量监测模型分别在蕾期、花期、铃期、吐絮期4个关键生育期对土壤氮含量的预测均达到了较高的精度,能够很好地反映棉花土壤氮素营养状况。利用植株冠层光谱参数可以很好地监测土壤氮素营养,说明利用植株冠层光谱方法监测土壤氮含量是可行的。     

基于红边参数的棉花冠层叶片氮积累量之估算研究

测试了棉花2个品种4水平种植密度的4个关键生育时期冠层反射光谱,应用微分技术处理棉花冠层反射光谱,提取了红边(680～750nm)波段范围的最大一阶微分值(Dr)和红边面积(SDr)参数。分析了棉花冠层红边参数在不同生育期的变化特征和棉花吐絮期的两种生长类型的冠层红边状况,表明红边位置可以指示它们的氮素状况。以新陆早8号的SDr为自变量与对应的LNA为因变量进行相关分析,SDr与冠层LNA达1%极显著相关(R=0.9186**,n=32),利用其构建的模型方程估算新陆早6号的LNA,实测LNA和估测LNA的估计标准差为0.8909g/m2,估算精度为88.1%(R=0.9277**,n=32),说明采用高光谱提取的红边参数信息是无损实时、快捷评价棉花氮素状况的有效方法。     

不同发育阶段‘文紫椒1号’果色色素的比例和组织定位

研究旨在阐明‘文紫椒1号’果实在不同发育阶段的色素比例和组织定位,为丘北辣椒在观赏园艺中的应用和观赏型新奇果色新品种的选育提供参考。以种植于云南文山州砚山县的‘文紫椒1号’果实为试材,将果实分为5个发育阶段,用分光光度法检测了果实的叶绿素含量([Chl])、类胡萝卜素含量([Car])、总黄酮类含量(TFC)、总花色苷含量(TAC)和辣椒红素含量([Cap]),并分析其配比和相关性,同时用徒手切片法检测了色素的组织定位。结果表明,在发育过程中,‘文紫椒1号’果实[Chla]/[Chlb]的最大值出现在发育阶段3,[Chl(a+b)]/[Car]、[Chl(a+b)]/TFC和[Chl(a+b)]/TAC的最大值均出现在阶段2,而[Chl(a+b)]/[Cap]、TFC/TAC、TFC/[Cap]和TAC/[Cap]的最大值均出现在阶段1。[Chla]、[Chlb]、[Chl(a+b)]、[Car]、TFC、TAC和[Cap]之间的相关性不尽一致,其中,TAC与[Chl(a+b)]、TFC之间均呈不显著正相关,而与[Car]之间呈不显著负相关,[Cap]则与[Chl(a+b)]、[Car]、TFC和TAC之间分别呈显著负相关、极显著正相关、不显著正相关和不显著负相关。果实色素主要定位在中果皮细胞中,细胞的色相和色度变化趋势与果色的高度一致。所以,‘文紫椒1号’的果色源于各发育阶段果皮细胞中叶绿素、类胡萝卜素（含辣椒红素）和黄酮类（含花色苷）色素的比例差异。     

棉花黄萎病相关基因GhAAT的克隆与功能鉴定

棉花黄萎病(Verticillium dahliae)是一种土传性真菌维管束病害,严重影响棉花的生产。本研究通过转录组数据分析筛选出一个与棉花黄萎病相关的氨基酸转运蛋白GhAAT基因,该基因在棉花根部响应黄萎病菌诱导表达。利用病毒诱导的基因沉默(virus-induced gene silencing, VIGS)技术研究其在棉花抗黄萎病中的功能。结果显示将该基因在棉花TM-1的叶片和根部沉默后,棉花对黄萎病抗性减弱,证明GhAAT基因参与调控了棉花抗黄萎病功能。发掘棉花抗黄萎病菌重要调控基因并揭示其分子机制对培育棉花抗病品种具有重要意义。     

Mechanism of Verticillium Wilt Stress Affecting Photosynthetic Characteristics and Chlorophyll Fluorescence Characteristics of Cotton Seedlings

[Objective] The goal of this experiment was to study the effects of Verticillium wilt on photosynthetic characteristics and chlorophyll fluorescence characteristics of cotton seedlings, and to provide a theoretical basis for the identification of cotton resistance to Verticillium wilt at the seedling stage and the establishment of Verticillium wilt control techniques. [Method] Zhongzhimian 2 (Verticillium wilt resistant cultivar, ZZM-2) and Jimian 11 (Verticillium wilt susceptible cultivar, JM-11) were selected. Verticillium dahliae was inoculated at the two true leaves stage. The photosynthetic and chlorophyll fluorescence parameters of the last fully expanded young leaves were measured at 6 d after inoculation. [Result] (1) Under the stress of Verticillium wilt, the leaf area per plant, leaf water potential, photosynthetic pigment content and net photosynthetic rate (Pn) of cotton seedlings decreased, and the decreases in ZZM-2 were smaller than those in JM-11. (2) The decreased intercellular CO2 concentration and increased stomatal limitation value in the two test materials indicated that the decrease in Pn was mainly caused by stomatal limitation, and the decrease in chlorophyll a in ZZM-2 was smaller than the decrease in chlorophyll b, whereas the opposite trend was observed in JM-11. (3) The light response curve parameters of ZZM-2 were increased and theCO₂ response curve parameters of ZZM-2 were decreased, but all parameters in JM-11 were decreased. For ZZM-2, the potential photochemical efficiency (F_v/F_m), the quantum yield of regulated energy dissipation (Y_NPQ) and the quantum yield of non-regulated energy dissipation (Y_NO) were increased slightly and the actual photochemical efficiency (Y_II) was decreased slightly, but the differences in these four indexes between diseased plants and healthy plants was not significant; however, the variable fluorescence yield (F_v) was significantly decreased. For JM-11, F_v, Fv/F_m, Y_II and Y_NPQ were significantly decreased, and Y_NO was significantly increased. (4) F_v/F_m in leaf lesion sites decreased, but that in non-lesion sites did not decline or even showed a slight increase. [Conclusion] Verticillium wilt resistant cultivars with higher photosynthetic capacity and carbon assimilation ability could reduce the effects of Verticillium wilt on photosynthetic characteristics and reduce the impacts on cotton yield and quality.     

Physiological Mechanism of Different Varieties and Potassium Application Amounts on Cotton Resistance to Verticillium Wilt

[Objective] It has theoretical guiding significance for the comprehensive control technology of Verticillium wilt of cotton to explore the effect of potassium nutrition on mechanism of cotton Verticillium wilt resistance under the condition of pathogen stress of Verticillium dahliae. [Method] Three cotton cultivars Jimian 11 (JM-11 as susceptible cultivar), Zhongzhimian 2 (ZZM-2 as control cultivar), Nongda 601 (ND-601 as resistant cultivar) were adopted, and each cultivar had 0, 75, 150, 225, 300 kg· hm－2 potassium levels represented by K0, K75, K150, K225, K300. Two factors split plot experiment was designed under artificial nursery inoculated Verticillium dahlia, to observe disease index of Verticillium wilt, and researched resistance associated substances content and enzyme activity. [Result] (1) The lignin, total phenol contents and POD, PAL activity in leaves or roots of JM-11 were more greatly influenced by Verticillium wilt compared with ZZM-2, ND-601; the crucial disease resistance substances and enzymes were lignin, total phenol and PAL for JM-11, total phenol for ZZM-2 and ND-601. (2) The lignin, total phenol contents and POD, PAL activities of JM-11 were more sensitive to root potassium than ND-601, but weaker than that of ZZM-2. The sensitivity of POD, PAL activities in functional leaves of JM-11 to potassium were more sensitive than ND-601 and ZZM-2. (3) The best effect of potash fertilizer for relieving disease index for JM-11, ZZM-2, ND-601 were 32.5%, 26.4%, 16.5%, and corresponding potassium application amounts were 290.5 kg·hm－2, 253.5 kg·hm－2 and 229.5 kg·hm－2, respectively. [Conclusion] Application of potassium could increase the resistance to Verticillium wilt and reduce the occurrence of cotton Verticillium wilt.     

陆地棉GhDAHP基因的克隆与表达分析

棉花黄萎病是制约棉花产业健康发展的一种重要病害,在生产上未发现有效的根治办法,使用现代基因工程技术,从基因库中挖掘抗黄萎病相关基因,为棉花抗黄萎病育种工作的提供理论基础。本研究根据在GenBank中检索到抗黄萎病基因SDAHP(GU479467,1,3-脱氧-阿拉伯庚酮糖-7-磷酸合成酶),从陆地棉基因组数据库中检索到3个同源基因(NM_016893351.1,XM_016849738.1和XM_0168151150.1),根据3个同源基因核苷酸序列设计引物,以陆地棉cDNA为模板进行克隆,获得3个目的基因,依次命名为GhDAHP-1、GhDAHP-2和GhDAHP-3。利用ProtParam和Prot Scale等在线软件对目的基因进行生物信息学分析;RT-PCR分析黄萎病菌诱导后目的基因在陆地棉叶片中的表达量。结果表明:GhDAHP-1序列全长1983 bp,氨基酸总数为539,分子质量约59.45378 kD,理论等电点pI为8.7;GhDAHP-2序列全长1832 bp,氨基酸总数516,分子质量约57.02998 kD,理论等电点pI为7.68;GhDAHP-3序列全长1652 bp,氨基酸总数517,分子质量约56.95491 kD,理论等电点pI为8.52。系统进化树分析,GhDAHP与雷蒙德氏棉(Gossypium raimindii)、木本棉(Gossypium arboreum)的亲缘关系最近。棉花叶片在黄萎病菌的处理下,GhDAHP基因表达量呈现先升高后降低的趋势,推测受到黄萎病诱导后,GhDAHP基因可能参与了黄萎病菌侵染的防卫过程。     

棉花品种对黄萎病慢病性初步研究

2002-2003年在河南省新乡县李台基地重病田,通过定株定时调查发病情况和各单株的产量结构,研究了不同抗病性棉花品种对黄萎病慢萎性。结果表明,若单纯以8月下旬的病情指数为标准,不少品种的抗病性均不理想,但从黄萎病发展历程分析,则发现有一部分耐至感病品种,其病情发展缓慢,对产量的影响也达不到显著水平。为此,提出棉花品种存在对黄萎病的慢病性,简称棉花品种慢萎性。     

棉花黄萎病抗性遗传和分子生物学研究进展

介绍了棉花黄萎病菌的致病机制 ,棉花黄萎病抗病机制及其遗传以及棉花黄萎病的鉴定方法 ,并对分子生物学在棉花黄萎病研究中的应用状况进展进行了综述。     

Induction of Defense Enzymes and Control Effect of Endogenous and Rhizosphere Bacteria on Verticillium Wilt of Cotton

[Objective] The colonization in soil, promoting cotton growth effects, inducing resistance of cotton and controlling effects to cotton Verticillium wilt of Bacillus vanillea SMT-24, B. velezensis BHZ-29, B. subtilis SHT-15 and B. atrophaeus SHZ-24 were studied in this paper, which provides a scientific basis for better control of cotton Verticillium wilt. [Method] The antagonistic resistance of these bacteria to Verticillium dahliae was judged by observing the inhibition zone; the number of colonization in the rhizosphere soil was tested to represent the colonization ability of antagonistic bacteria; the growth promoting effect of strains on cotton was determined by analyzing cotton plant height, root length, root hair number and number of leaves; the effect of antagonistic strains on cotton induced resistance was explored by determining catalase (CAT), superoxide dismutase (SOD), phenylalanine ammonia lyase (PAL), polyphenol oxidase (PPO), and peroxidase (POD) activities in cotton leaves, and investigating the disease index of cotton after inoculation with antagonistic bacteria solutions. [Result] SMT-24, BHZ-29, SHT-15 and SHZ-24 strains significantly inhibited the growth of V. dahliae and formed a clear zone of inhibition. After 2 to 22 days of the third inoculation, these strains of bacteria could survive in the root soil, increase the plant height, leaf number, and root hair number of cotton plants, as well as the activities of cotton CAT, SOD, PAL and POD enzymes, but reduced the disease index. [Conclusion] SMT-24, BHZ-29, SHT-15 and SHZ-24 strains can promote cotton growth, increase the activities of some related defense enzymes, and effectively control cotton Verticillium wilt.     

Hyperspectral Models for Estimating SPAD Value of Cotton Leaves under Waterlogging Stress

[Object] To setup the hyperspectral sensing models for estimating SPAD value of cotton leaves under waterlogging stress. [Method] Irrigation and drainage controllable plots were introduced to simulate the waterlogging stress treatment in the flowering and boll forming stage, during which the change characteristics of the cotton leaf spectral reflectance and SPAD value were observed after 1 d, 3 d, 6 d, 9 d waterlogging, respectively. To find out the hyperspectral sensing models for estimating SPAD value of cotton leaves under waterlogging stress, the correlation and regression relationships between SPAD value and spectrum parameters were analyzed. [Result] (1) The SPAD value of the fourth cotton leaf from the top was significantly lower than control when suffers from waterlogging for 3 d, when waterlogged 9 d the SPAD value decreased by around 15% compared with the control. (2) The cotton suffering from waterlogged damage in the flowering and boll forming stage caused the reflection peak in green light wave band became steep, while the near infrared spectral reflectance increased, and caused the reduction of red absorption and red edge position "blue shifts", the red edge position drifts towards short wave with 4～5 nm when suffers from waterlogging for 9 d. With increase of the waterlogged days, the red edge slope and red edge area increased with a maximum value at 6 d of waterlogging, meanwhile, the skewness and kurtosis of red edge increased. (3) After waterlogging, the SPAD value of the fourth cotton leaf from the top (chlorophyll content) had a remarkable correlation with red edge slope(Dr), red edge position(λr), green peak reflection(Rg), green peak position(λg), red well position(λo), blue edge area(SDb), yellow edge skewness(Sy), yellow edge kurtosis(Ky), red edge skewness(Sr), red edge kurtosis(Kr), etc. An experience linear, polynomial and exponential models for estimating SPAD value had been built through using the Sy, Sr, Kr as independent variables, respectively, their determination coefficient (R²) were greater than 0.9, and the root mean square error (RMSE) were less than 1; and an experience binary linear regression equation for estimating SPAD value had been built through multivariate regression using the λg, SDr/SDb(VI3), Sb, Sy, Ky as independent variables, the R² was as high as 0.973, and the RMSE was 0.393. [Conclusion] The model can be remote sensing model used as estimating leaf SPAD of cotton value under waterlogging stress.     

短季棉花黄萎病田间分布型初步研究

通过对短季棉重病田的研究表明，棉花黄萎病株在田间呈正态分布，对棉株产量影响大的重病株则呈负正态分布，而几乎绝产的枯死病株也呈负二项分布，说明危害棉花产量大的病株在田间呈聚集状态。