田菁茎瘤固氮根瘤菌在小麦体内的定殖及营养元素相关miRNA的表达

【目的】随着化肥过度使用引起的环境问题的出现,无公害农业的推广,以及新型肥料研究领域不断拓宽,微生物肥料,尤其是植物根际促生菌的研究成为近年来的热点。然而微生物肥料的增产机理还基本停留在作物农学性状的表观调查上,没有从分子水平进行研究。因此,本文用田菁茎瘤固氮根瘤菌（Azorhizobium caulinodans ORS571）侵染小麦,探索GFP-A.caulinodans在小麦幼苗组织中的分布与定殖规律以及营养元素相关miRNAs在小麦与A.caulinodans互作中的作用机制。【方法】使用A.caulinodans侵染小麦种子（品种为小偃22）,将接菌6 d后的小麦幼苗的根和接菌12 d后的叶制作玻片,利用激光共聚焦显微镜对样品进行逐层扫描,检测GFP-A.caulinodans在幼苗不同组织中的分布与定殖情况。同时,采集接菌后0 h、 6 h、 12 h、 24 h、 48 h、 72 h、 96 h的小麦幼根取样,用Trizol法提取总RNA。利用试剂盒进行加尾和反转录反应,将样品总RNA中的miRNA合成为cDNA,使用-tubulin作为内参基因,进行实时定量PCR反应,使用2-CT方法计算相对表达量。利用psRNATarget 在线软件,采用默认参数,对miRNA的靶基因进行预测。【结果】 1）激光共聚焦结果显示,GFP-A.caulinodans可定殖于根的表皮细胞、 细胞间隙、 根尖破损处和根毛,在根维管组织和叶片气孔部位,也发现有GFP-A.caulinodans存在。2）实时定量PCR分析结果表明,6条与营养元素代谢有关的miRNA表达发生变化,其中miR164、 miR167和miR827相对表达量呈现出先上调后下调的趋势,miR169 和miR398相对表达量也基本呈现出这一趋势。miR164、 miR167、 miR169和miR398的相对表达量在接菌12 h时上调至最高点,分别为对照的4.13、 2.84、 2.46和3.99倍; miR827相对表达量在接菌24 h时达到最高点,为对照的2.17倍。miR399相对表达量呈现出先下调后上调的趋势,在接菌24 h时降至最低点,为对照的0~21倍。3）通过靶基因预测,6条miRNA的靶基因分别编码了NAC1转录因子、 生长素响应因子、 HAP转录因子、 Cu/Zn 超氧化物歧化酶、 蛋白缀合酶PHO2和SPX-MFS亚家族蛋白。【结论】GFP-A.caulinodans能够从根毛和根尖破损处等部位进入小麦幼苗根部定殖,并可通过向上迁移到达叶片,在气孔处定殖。接种A.caulinodans可不同程度增加小麦根中响应氮素、 磷素、 微量元素的miRNAs相对表达量,增强小麦幼苗对营养元素的吸收和利用,促进小麦根的形态建成。     

钩状木霉 ACCC31649的GFP标记及其对辣椒定殖和促生作用

【目的】旨在建立农杆菌介导的绿色荧光蛋白 (GFP) 基因转化钩状木霉 (Trichoderma hamatum) ACCC31649的技术方法,筛选遗传稳定的GFP标记转化子,并研究该菌株在辣椒植株中定殖和对辣椒的促生作用,为进一步阐明木霉在辣椒根际定殖及其与辣椒病原菌在辣椒根际和植株中的定殖、互作和生防作用奠定基础。【方法】通过根癌农杆菌介导的遗传转化方法筛选遗传稳定的GFP标记转化子,通过灌根接种方法和组织切片的水玻片荧光观察研究了钩状木霉在辣椒植株中定殖过程和对辣椒的促生作用。【结果】获得了遗传稳定GFP标记的钩状木霉转化子。荧光显微观察表明,辣椒根、茎、叶组织中都检测到GFP标记菌株的定殖。标记菌株首先在根部定殖,然后通过根部维管束逐步定殖到茎和叶片组织中。野生型菌株和GFP标记菌株灌根接种4叶期辣椒幼苗,30天后,GFP标记菌株与水处理对照相比,辣椒的株高增长13.5%,根长增长16.2%,鲜重和干重分别增加了43.8%和45.3%,而且野生型菌株与GFP标记菌株对辣椒的促生作用没有显著差异。【结论】钩状木霉能够在辣椒植株根、茎和叶组织中定殖,并且对辣椒具有显著的促生作用。同时,GFP标记的钩状木霉将在进一步阐明该菌株在辣椒根际定殖及其对病原菌拮抗和互作研究中发挥重要作用。     

甘蔗内生菌B9的鉴定及其促生长机制和定殖能力的研究

B9是一株分离于甘蔗品种"云蔗99-91"根部的内生菌,笔者对其种属鉴定与促生长机制开展研究,以期应用于甘蔗生产。试验通过传统方法与16S rRNA等看家基因片段测序对菌株进行分类鉴定;采用固体透明圈、酸性钼蓝比色法、火焰分光光度法、GFP荧光标记等方法测定与研究B9菌株固氮、溶磷、解钾、产IAA、嗜铁素等生物学活性及在甘蔗苗内的定殖情况;最后通过回接法研究菌液处理对玉米种子萌发与甘蔗幼苗生长发育的影响。结果表明B9菌属于芽孢杆菌属枯草芽孢杆菌(Bacillus subtilis),具有固氮、溶磷、解钾、产生IAA和嗜铁素的能力。用B9菌液处理后能提高玉米种子的发芽势与促进根芽组织生长,同时处理组甘蔗苗株在农艺性状与生理生化水平上明显优于无菌清水对照,并且发现B9菌能在甘蔗根茎叶内稳定定殖。本文鉴定了B9菌的种类和功能,阐明了B9菌的促生长机制和定殖能力,为开发生物菌肥并应用于生产提供理论依据。     

缩二脲降解菌在作物根际的GFP标记示踪

通过三亲结合对缩二脲降解菌GW-1菌株成功进行了绿色荧光蛋白（Green fluorescent protein, GFP）基因标记,标记菌株命名为GW-1-GFP。实验证明外源质粒对宿主菌的生长未带来不利的影响,且经高效液相色谱测定该标记菌株降解缩二脲的能力与出发菌株无显著差异（p<0.05）。经抗生素抗性和荧光追踪证明标记菌株GW-1-GFP能够很好地在土壤中定殖,第25天时在不同处理的土壤中标记菌株对缩二脲的降解能力都超过50%,45天后该标记菌株在土壤中已难以检测。经标记菌株GW-1-GFP菌悬液浸润的小麦种子发芽后,通过荧光观察显示降解缩二脲的标记菌株GW-1-GFP在小麦根部定殖良好,且该标记菌株能在一定程度上缓解缩二脲对小麦的毒害作用。该研究为验证、追踪土壤中功能微生物菌剂与作物根部的亲和性和生态有效性提供了简易、直观的检测方法。     

黄淮麦区推广品种小偃22农杆菌遗传转化体系的建立

小偃22小麦是目前陕西省生产上的主栽品种,在黄淮麦区其他地区也大面积推广种植。本文通过对小偃22小麦胚性愈伤组织的来源﹑生理状态和干燥时间,根癌农杆菌的接种浓度和侵染方式,AS的添加浓度,胚性愈伤组织和根癌农杆菌的共培养温度和共培养时间,潮霉素的筛选浓度和筛选方式等参数进行比较研究,建立了小偃22小麦农杆菌遗传转化体系。结果显示,以来源于幼穗的胚性愈伤组织作为转化受体较为适宜,愈伤组织生理状态为从幼穗直接诱导30d后继代1次并培养15d的胚性愈伤组织,农杆菌侵染前对胚性愈伤组织干燥处理30 min较为适宜;农杆菌菌液浓度为0.8OD,采用抽真空侵染5min＋非真空侵染10min的浸染方式,共培养温度为22℃,共培养时间为72h,AS的适宜添加浓度为150μmol/L;筛选方式选用在抗性愈伤组织筛选阶段经75mg/L潮霉素筛选后,在分化阶段不再进行筛选或采用25mg/L低浓度潮霉素筛选。通过重复转化验证试验和分子生物学鉴定,表明该遗传转化体系具有可行性。     

内生菌YN201728的定殖能力及其防治烟草白粉病的效果研究

为研究生防菌YN201728在烟草体内的定殖规律及防病机制,本研究利用其绿色荧光标记菌株YN28-P43GFPmut3a实时动态观测标记菌的定殖部位及密度,并探究其对温室烟草白粉病的盆栽防效与定殖的关系。结果表明,YN28-P43GFPmut3a发酵液处理烟草种子和幼苗后,种子内的标记菌含量可达2.18×10⁶ CFU·g^-1,在幼苗的根表土、根际土、根、茎、叶等组织中均能检测到标记菌,且其定殖密度表现为根表土>根际土>根>茎>叶。激光共聚焦显微镜观察显示,标记菌主要聚集在烟草根表皮、木质部导管、茎表皮、韧皮部及维管束组织、叶片表面和叶肉细胞间隙以及种皮与胚等部位。盆栽防效试验结果表明,YN201728野生型和标记菌株对烟草白粉病均有较好的保护和治疗效果,持效期长达21 d。此外,烟草叶片中内生菌的定殖量与其防效呈正相关。本研究结果表明,内生菌YN201728在烟草体内有良好的定殖和防治白粉病效果,具很好的开发潜力,为烟草病害的生物防治提供了一定的理论基础。     

巨大芽胞杆菌WY4的GFP标记及其在小白菜上的定殖

解磷菌(phosphate solubilizing bacteria,PSB)可以通过提高土壤有效磷含量而增加作物产量,目前已有许多解磷菌被分离并应用于农业生产中,但关于解磷菌在植物根际中的定殖情况仍缺乏系统性的研究.WY4为本实验室前期从小白菜(Brassica chinensis)根际分离得到的一株高效解磷菌,本研究利用绿色荧光蛋白(green fluorescent protein,GFP)标记技术研究了WY4在小白菜根际及土壤中的定殖规律.与原菌株相比,GFP标记对菌株WY4-GFP生长及解磷活性具有较小影响,同时在促进小白菜生长上WY4-GFP与WY4无显著性差异;WY4-GFP具有持久的定殖能力(接种21d的自然土及30 d的小白菜根际土中,WY4-GFP的定殖数量分别为105和104 CFU/g左右),同时随时间的增加WY4-GFP在土壤中定殖数量逐渐减少;WY4-GFP在小白菜根冠及分生区大量定殖,在伸长区及侧根根毛处数量较少,同时表皮细胞间隙上也有较多的标记菌株.研究结果表明,WY4-GFP在小白菜根际及土壤中具有良好的定殖能力,这为后期深入研究解磷菌与植物间的关系提供了重要参考.     

解磷细菌 K3 的 GFP 标记及其解磷能力检测

本文运用构建成功的含有假单胞菌属自身启动子及绿色荧光蛋白(GFP)的质粒pTRGFP电转至假单胞菌解磷细菌K3中,通过激光共聚焦显微镜及质粒检测,获得了发光稳定的标记菌株K3GFP。7天液体摇瓶试验中,标记菌株K3GFP的可溶性P含量在第4天达到最高774μg/ml,而出发菌株K3在第3天时已经达到了最大含量780μg/ml,说明pTRGFP质粒的转入对K3菌株解P能力有一定的影响。标记菌株K3GFP施入自然土壤10天后数量维持在5.47×106CFU/g～2.40×106CFU/g,35天后降到5.0×103CFU/g,说明解磷细菌K3GFP可以在自然土壤中定殖。本试验为研究解磷细菌在根际及土壤中的生长动态等行为特征奠定了基础。     

解淀粉芽孢杆菌41B-1R对棉花黄萎病的防效研究

棉花黄萎病严重危害棉花生产,尚无有效的控制方法。为提高棉花对黄萎病的抗性,本研究将从棉花根系中分离的对棉花黄萎病菌有较好抗性的内生细菌解淀粉芽孢杆菌41B-1R菌株添加到育苗基质中,培育苏棉22和中棉所41 2个棉花品种的幼苗,并评价生物育苗基质对棉花生长及抗黄萎病的效应。结果表明,育苗基质适合作为41B-1R的保存载体;添加生防菌41B-1R的育苗基质对棉花幼苗生长无不良影响;41B-1R能稳定地定殖于棉花幼苗根系内部。育苗后移栽,可有效减少非落叶型棉花黄萎病菌V1070在苏棉22和中棉所41幼苗根部的定殖,显著提高棉苗对黄萎病的抗性,防治效果分别达到46.7%和28.6%。本研究表明,用添加41B-1R菌株的育苗基质育苗可以有效降低棉花黄萎病的危害,为棉花黄萎病的防治提供了新途径。     

不同浓度外源-氧化氮对盐胁迫下小麦幼苗生理特性的影响

为探讨添加不同浓度外源一氧化氮(Nitric oxide,NO)对小麦盐害的缓解作用,确定最适添加浓度,本研究以山农22号为供试小麦品种,以硝普钠(Sodium-nitroprusside,SNP)为外源NO供体,采用液培方式,研究不同浓度SNP(0、25、50、100、150、200μmol L-1)对100 mmol L~(-1)NaCl胁迫下小麦幼苗生长及生理特性的影响。结果表明,100 mmol L~(-1)NaCl显著抑制了小麦幼苗的生长,添加不同浓度SNP均能缓解盐胁迫对小麦幼苗生长的抑制作用,提高小麦幼苗体内抗坏血酸(Ascorbic acid,As A)含量和亚细胞各组分中超氧化物歧化酶(Superoxide dismutase,SOD)活性,减少丙二醛(Malondialdehyde,MDA)和过氧化氢(Hydrogen peroxide,H_2O_2)的积累,减轻膜脂过氧化毒害;促进脯氨酸的合成与积累,降低电解质渗透率;抑制小麦对Na的吸收,增加对其它必需矿质养分的吸收;提高小麦幼苗叶片的叶绿素含量,促进光合作用;增加小麦幼苗的干鲜重,促进小麦生长。不同浓度SNP对小麦幼苗盐胁迫的缓解作用随SNP浓度的增加先升高后降低,以100μmol L~(-1)SNP对小麦幼苗盐胁迫的缓解作用最为明显。     

小麦与油菜种子萌发对酸雨胁迫的反应研究

用pH2.0、2.5、3.0、3.5、4.0、5.0模拟酸雨处理小麦和油菜种子试验结果表明,pH2.0时小麦和油菜种子不发芽,pH2.5时只有小麦异状发芽,pH≥3.0时小麦和油菜种子发芽率、发芽势、发芽指数、活力指数均与pH值显著正相关,小麦异状发芽率则随pH值上升而降低。小麦和油菜的吸水值、呼吸速率、贮藏物质运转效率也与pH值显著正相关,小麦贮藏物质消耗率与pH值显著正相关,而油菜与pH值显著负相关;小麦和油菜的根、芽长抑制指数均与pH值显著负相关,且小麦抗酸雨胁迫能力强于油菜。     

Root architectural responses of wheat cultivars to localised phosphorus application are phenotypically similar

Differences in nutrient recovery from fertiliser bands may improve cereal variety selection. The objective of this study was to identify the variation in root plasticity across commonly grown Australian wheat (Triticum aestivum L.) cultivars in response to a phosphorus (P)‐enriched band. Ten wheat cultivars were screened for root proliferation within a 150 mg P kg⁻¹ band in P‐responsive soil. Plants were destructively harvested at the four‐leaf phenological stage and various growth parameters, including root length density (RLD), were measured on banded and uniformly adequate P treatments. All wheat cultivars increased RLD between three and nine times in the P band. However, there was no significant difference in root plasticity among the cultivars tested. Although all cultivars produced longer, though ≈ 9% thinner roots when responding to the P band, the phenotypic response was unable to compensate fully for the lower P status encountered in the soil. Despite 23% longer root lengths in the P‐band treatments, P uptake per unit root length was 78% lower than in uniformly adequate P treatments. Our results indicate that root plasticity of wheat cultivars in a P‐enriched band was phenotypically similar. Further research is necessary before selecting for wheat cultivars that respond to localised nutrient patches with increased RLD.     

小麦苗期氮素吸收利用效率差异及聚类分析

【目的】氮肥过量施用,不仅造成氮素大量流失,还增加了农业生产成本,对生态环境带来了巨大的威胁。筛选和培育氮高效小麦品种是提高氮肥利用率、 降低环境污染风险的有效途径。本文通过对44个小麦品种苗期性状的考察,初步筛选出具有氮高效潜力的小麦品种。【方法】利用循环营养液培养方法,研究了安徽省44个小麦品种(系)在正常氮(5 mmol/L)和高氮(45 mmol/L)条件下苗期氮素吸收利用效率的差异。采用隶属函数法将评价指标数据进行标准化,区间为[0,1];而后采用客观赋权法将标准化后的数据整合成一个无量纲的综合值,最后基于综合值运用最短距离法、 欧氏距离平方聚类分析方法,将44个小麦品种划分成不同的氮效率类型。【结果】在两种供氮水平下,不同小麦品种的茎叶干重、 根干重、 叶面积、 茎叶氮累积量和根氮累积量存在显著性差异,其变异系数分别在27.9%～33.7%和21.5%～32.8%之间,可作为小麦苗期氮效率的评价指标。小麦苗期氮效率综合值在正常氮和高氮水平下分别在0.053～0.920和0.001～0.853之间,其中鉴76在正常氮和高氮条件下的氮效率综合值均大于80%。通过隶属函数氮效率综合值及其聚类分析,将44个供试小麦品种分为氮高效型、 氮中效型和氮低效型三类;其中扬麦16和鉴76在正常氮和高氮条件下均表现为高效型,皖麦68、 F60501-4、 鉴62和安农1026只在高氮条件下表现为高效型。氮高效型、 氮中效型、 氮低效型小麦品种在正常供氮和高氮条件下分别占供试品种总数的4.54%、 54.55%、 40.91%和13.63%、 38.64、 47.73%。【结论】在正常供氮和高氮条件下,44个供试小麦品种的茎叶氮累积量、 茎叶干重、 根部氮累积量、 根部干重和叶面积存在显著性差异,可以作为小麦苗期氮效率评价指标;初步确定扬麦16和鉴76为正常供氮和高氮条件下的氮高效型品种,皖麦68、 F60501-4、 鉴62和安农1026 为高氮条件下的氮高效型品种。     

Pseudomonads as a factor in the growth of winter wheat (Triticum aestivum L.)

Pseudomonads were isolated from the roots of winter wheat plants. Plants produced from direct-drilling of seeds into plots where crop residues had been burnt were larger and appeared healthier than those from plots where crop residues remained. Tests showed that roots from the plants direct-drilled into the plots where crop residues had been burnt were colonized by fewer pseudomonads which could inhibit plant growth than those from plants produced from plots where crop residues remained. The tests were performed by inoculating wheat seedlings with individual bacterial isolates and measuring root growth. When several wheat cultivars were bioassayed against each of two of the inhibitory pseudomonads, they differed greatly in susceptibility to the adverse effects of the bacteria.     

两个小麦品种对根际土壤中磷的吸收

Inorganic soil phosphorus extractable with sodium bicarbonate(NaHCO3-Pi),soil pH and root hairs length and density in the rhizosphere of two winter wheat cultivars (Tritium aestivum L.cv.Shichum,Sleipner)grown on a high pH Chinese silt loam(52.7 mg NaHCO3-Pikg^-1) and a Danish sandy loam(43.4mg NaHCO3-Pi kg^-1)wer studied to assess how these wheat cultivars differed in phosphorus uptake.The rhizosphere soil pH of two wheat cultivars grown on the two soils were fairly unchanged with increasing distrance from the roo surface.However the root hairs of Shichun were 2.1 times longer than those of Sleipner,Root surface area(RSA) of Shichun increased by 192% due to root hairs whereas root hairs of Sleipner increased RSA by 68% only.Hence the root system of Shichun was in contact with more soil than that of Sleipner,even though Sleipner had a longer root,Grown at the lower pH and level of NaHCO3-Pi in the Danish soil Shichun absorbed more inorganic phosphorus than Sleipner whereas at the higher pH and level of NaHCO3-Pi in the Chinese soil there was no phosphorus uptake difference between the two wheat culivars.     

Growth stimulation and nitrogen supply to wheat plants inoculated with Azospirillum brasilense

Twelve Azospirillum brasilense strains isolated from wheat (Triticum aestivum L.) roots were compared for root colonization, growth stimulation, and nitrogen (N) supply to young wheat plants cv. Klein Chamaco grown in sterile nutrient solutions without N. All the strains inoculated colonized both the root surface and interior, and most strains stimulated root and shoot growth, although the degree of stimulation was different for the different strains. Some strains increased the total N content of roots and tops at the end of the experiment, in one case up to 80% of the uninoculated plants, while others produced no effect on N content. No correlation could be found between growth stimulation or the amount of N supplied to the plant with the degree of root colonization. When the most efficient strain for N fixation was inoculated to different wheat cultivars, it stimulated growth and supplied N to the five cultivars tested, although the degree of root colonization, growth stimulation and N supply showed differences among the cultivars. Our results suggest that there exists the potential of A. brasilense to supply N to wheat plants in considerable amounts, although an adequate strain are still to be identified.     

接种食细菌线虫对小麦生长和N、P吸收的影响

A 40-day gnotobiotic microcosm experiment was carried out to quantify the effect of bacterial-feeding nematode on plant growth and nutrient absorption. The results showed that inoculation of bacterial-feeding nematode Protorhabditis sp. stimulated the growth of wheat (Triticum aestivum) and the uptake of N. By the end of the 40-day incubation wheat biomass and N uptake in the treatment with nematode and bacteria (Pseudomonas sp.) increased by 6.5% and 5.9%, respectively, compared with bacteria alone treatment. The presence of nematode mainly accelerated the growth of aboveground of wheat, while it slightly inhibited the root development. There was little difference in plant tissue N concentration between treatments. P concentration and uptake of wheat, however, were generally reduced by nematode. It appears that the enhancement of plant growth and nitrogen uptake is attributed to the enhancement of nitrogen mineralization induced by nematode feeding on bacteria, and the reduction of phosphorous uptake is the result of weak root status and competition by bacteria immobilization.     

Biostimulant effects of Bacillus strains on wheat from in vitro towards field conditions are modulated by nitrogen supply

Bacillus velezensis strains, belonging to plant growth‐promoting rhizobacteria (PGPR), are increasingly used as microbial biostimulant. However, their field application to winter wheat under temperate climate remains poorly documented. Therefore, three B. velezensis strains IT45, FZB24 and FZB42 were tested for their efficacy under these conditions. Two biological interaction systems were firstly developed under gnotobiotic and greenhouse conditions combined with sterile or non‐sterile soil, respectively, and finally assayed in the field during two years coupled with different N fertilization rates. Under gnotobiotic conditions, all three strains significantly increased root growth of 14 d‐old spring and winter wheat seedlings. In the greenhouse using non‐sterile soil, only FZB24 significantly increased root biomass of spring wheat (+31%). The three strains were able to improve nutrient uptake of the spring wheat grown in the greenhouse, particularly for the micronutrients Fe, Mn, Zn, and Cu, but the observed increases in nutrient uptake were dependent on the organs and the elements. The root biomass increases in inoculated plants coincided with lowered nutrient concentrations of P and K. In 2014, under field conditions and absence of any N fertilizer supply, FZB24 significantly increased grain yields by 983 kg ha^?1, or 14.9%, in relation to non‐inoculated controls. The three strains in the 2015 field trial failed to confirm the previous positive results, likely due to the low temperatures occurring during and after inoculations. The Zeleny sedimentation value, indicative of flour quality, was unaffected by the inoculants. The results are discussed in the perspective of bacterial application to wheat under temperate agricultural practices.     

Natural Fluorescence of Red and White Wheat Kernels

Red and white wheats must be segregated for marketing purposes because they have different end uses. Identification of wheat color is not straightforward, and currently there is interest in characterizing red and white wheats using spectroscopic methods and chemical tests. The kernels of both red and white wheats exhibit natural fluorescence that can be readily viewed under UV light, although it is not possible to differentiate the fluorescence spectra of red and white wheats by visual inspection only. Fluorescence emission spectra in the wavelength range of 370–670 nm for 91 wheat samples consisting of 48 red (from 30 cultivars) and 43 white (from 18 cultivars) were analyzed by partial least squares (PLS) and neural networks analyses (NNA). Samples included cultivars that were difficult to classify visually as well as wheat harvested after rainfall. Classification accuracies were ≈85% for calibration and ≈72% for the validation samples by both analyses. A plot of β‐coefficient vs. wavelength in PLS analysis indicated that fluorescence of red wheat cultivars was greater than that for white wheat cultivars at 425 (±20) nm wavelength. Fluorescence of white wheat cultivars was greater than that for red cultivars at 587 (±35) nm. Fluorescence emission at ≈450 nm from wheat samples increased in intensity after treatment with NaOH. The increase was greater for red than for white wheat. Wheat harvested after rainfall also exhibited a slight increase in fluorescence.