不同光照和水分下三叶鬼针草与本地种金盏银盘生长特征的比较研究

为探讨三叶鬼针草(Bidens pilosa)成功入侵机制,利用盆栽试验对不同光照和水分条件下三叶鬼针草与本地种金盏银盘(B.biternata)的生长特征进行了比较研究。结果表明,两物种对光和水的变化均具有较大可塑性,但与金盏银盘相比,在有利环境下三叶鬼针草具有较大的株高、叶面积、生物量;在不利环境下,三叶鬼针草具有较大的比叶面积和叶面积比;除叶面积比和叶生物量比外,三叶鬼针草各生理参数的可塑性指数均高于本地种。这说明三叶鬼针草具有较强的表型可塑性和入侵适应性,这些特性促进了其成功入侵。     

Ecological significance of root tip rotation for seedling establishment of Oryza sativa L

How plant seeds secure root penetration into soil to obtain good seedling establishment is one of the basic ecological problems. In this study, seminal root growth was investigated to clarify the cause of varietal difference of seedling establishment in direct seeding of rice in flooded paddy fields, with special reference to root tip rotation. In a field experiment, seedling establishment percentage had a weak correlation with seminal root elongation rate but was not correlated with apparent seedling weight in water, which has been reported to be the cause of floating seedlings resulting in poor seedling establishment. Root tip rotation was analyzed for indoor-grown seedlings using spectrum analysis: the maximum entropy method (MEM) was used. Maximum entropy method power spectrum analysis clarified that maximum MEM power density (practically corresponds to spiral angle) detected in the frequency range above 0.1 cycles mm^-1 was highly and positively correlated to seedling establishment percentage in the field experiment. Maximum MEM power density in high correlation with seedling establishment was mostly found around frequencies of 0.2 cycles mm^–1, which corresponded to 2.0–3.4 cycles of root tip rotation per day. From these results, root tip rotation (circumnutation) with a larger spiral angle was suggested to play an important role in the establishment of rice seedlings on flooded and very soft soil. A possible explanation for why a larger spiral angle was advantageous for seedling establishment is that if buoyancy and seedling weight are constant, a larger pushing force of the seminal root is available without causing floating of a seedling, due to the upward force being a reaction of the seminal root pushing force.     

Nitrate levels affect the development of the black locust-Rhizobium symbiosis

Summary Experiments with black locust (Robinia pseudoacacia L.) seedlings grown under strictly controlled laboratory conditions indicated that the availability of nitrate has a marked impact on nitrogen fixation. When nitrate concentrations were very low, both nodulation and seedling growth were impaired, whereas nitrate concentrations high enough to promote plant growth strongly inhibited symbiotic nitrogen fixation. When nitrate was added to the growth medium after infection, nodulation and nitrogen fixation of the seedlings decreased. This effect was even more marked when nitrate was applied before infection with rhizobia. Higher nitrogen concentrations also reduced nodule number and nodule mass when applied simultaneously with the infecting bacteria. The contribution of symbiotic nitrogen fixation to black locust shoot mass by far exceeded its effects on shoot length and root mass. When nitrate availability was very low, specific nitrogen fixation (i. e. nitrogenase activity per nodule wet weight) was improved with increasing nitrogen supply, but rapidly decreased with higher nitrogen concentrations.     

Different responses to shade of evergreen and deciduous oak seedlings and the effect of acorn size

An evergreen oak species, Cyclobalanopsis multinervis, and a deciduous oak species, Quercus aliena var. acuteserrata were grown from acorns under two light levels (full sunlight and shade at about 18 % of full sunlight, simulating the light intensities in forest clearings and gaps, respectively) for one growing season. Three hypotheses were tested: (i) the deciduous species grows faster than the evergreen species in forest gaps and clearings; (ii) the deciduous species responds more strongly in terms of growth and morphology to variation in light climate than the evergreen species; and (iii) seedling size is positively correlated to acorn size. The results showed: (i) at both light levels, the deciduous seedlings gained significantly more growth in biomass and height than the evergreen seedlings; (ii) both species produced significantly more biomass in full sunlight than in shade, without showing any significant difference in height between treatments. Increase in light intensity improved the growth of the deciduous seedlings more strongly; (iii) at a similar age, the deciduous seedlings showed a greater response in leaf morphology and biomass allocation to variation in light levels, but when compared at a similar size, biomass allocation patterns did not differ significantly between species; (iv) bigger acorns tended to produce larger seedlings, larger leaf sizes and more leaf area, between and within species. These differences demonstrate that the deciduous species is gap-dependent and has the advantage over the evergreen species in forest gaps and clearings.     

附生植物石斛的株丛生长及营养繁殖特征

石斛(Dendrobium nobile Lindl.)为兰科多年生附生性草本植物,特化的假鳞茎是其营养贮藏器官,通过假鳞茎可实现克隆生长。该研究以野外调查发现的石斛株丛为研究材料,比较不同等级株丛假鳞茎合轴生长和高位腋芽的差异,分析高位株丛的定植方式,探讨石斛株丛生长及营养繁殖对附生环境的适应策略。结果显示:(1)石斛株丛的生长和扩大通过合轴生长的营养繁殖方式进行,假鳞茎基部具有2~3个储备芽,每年萌发1~2个新芽,随着生长年限的增加,形成大小不一的株丛。(2)株丛具有典型的高位腋芽营养繁殖特性,且主要形成于假鳞茎密集和老根密布的大株丛。(3)高位株丛母茎一端附着于附主树种上,在母茎软化和高位株丛的重力作用下,缩短了高位株丛与附主的距离,使其根系能够触及附主,完成高位株丛的定植。研究表明,附生植物石斛通过假鳞茎合轴生长的营养繁殖方式来增强并延续株丛寿命,高位腋芽的频发是株丛假鳞茎对拥挤等逆境的响应,高位株丛的定植依赖于母茎,这也是石斛对附生环境的一种生态适应策略。     

摩西球囊霉对三叶鬼针草保护酶活性的影响

旱地农田入侵杂草三叶鬼针草(Bidens pilosa L.)与摩西球囊霉(Glomus mosseae)(AM真菌)经常形成长效的共生体,该霉菌对三叶鬼针草的入侵能力起到促进作用,但机理并不清楚。盆栽试验对正常浇水、中度干旱和重度干旱条件下接种AM真菌的三叶鬼针草植株与未接种植株之间叶片丙二醛(MDA)含量及超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、抗坏血酸氧化酶(ASP)和过氧化物酶(POD)等保护酶活性进行了比较研究。结果表明,干旱胁迫导致三叶鬼针草叶片内MDA含量升高,SOD、CAT、ASP和POD的活性升高;正常浇水条件下,接种G. mosseae 对MDA含量,SOD、ASP和CAT活性影响不显著;中度干旱条件下,接种没有显著影响ASP活性,但对SOD和CAT活性影响显著;在处理前期(7,14,21d)POD活性影响不显著,在处理后期(28,35d)接种植株显著低于未接种植株;重度干旱条件下,未接种植株MDA含量、CAT活性显著高于接种植株,POD活性差异不显著。ASP活性在21d前差异不显著,之后,未接种植株显著高于接种植株。因此,AM真菌G. mosseae 有效地降低了干旱胁迫对三叶鬼针草的伤害程度,随着土壤含水量的严重亏缺和胁迫时间的延长,摩西球囊霉对三叶鬼针草的保护作用逐渐减弱。由于三叶鬼针草和AM真菌之间普遍存在着共生关系,该共生关系可能是三叶鬼针草入侵能力强的关键生物因子之一。     

Calcium and calmodulin inhibitor effects on the growth of carrot (Daucus carota L.) and radish (Raphanus sativus L.) in nutrient culture

Growth of carrot and radish seedlings in nutrient culture was inhibited by pretreatment with three calmodulin inhibitors. There was little selective effect on specific organs, shoots, tap root and fibrous roots over a range of concentrations. Although pretreatment with CaCl₂ (0.5 mM) did not affect growth of untreated seedlings, it partially reduced the inhibitory effects of trifluoperazine over the concentration range 0.01–0.05 mM. Trifluoperazine reduced the growth of GA₃-treated seedlings but did not overcome the modifying effect of GA₃ in favouring shoot/root ratio; ABA exacerbated its inhibitory effect on overall seedling growth and particularly on tap root development.Abbreviations GA₃ gibberellic acid - ABA abscisic acid - CaCl₂ calcium chloride - GAs gibberellins - Tfp trifluoperazine     

Lazy gene (la) responsible for both an agravitropism of seedlings and lazy habit of tiller growth in rice (Oryza sativa L.)

Using an isogenic line of rice having lazy gene (la), we studied the correlation between the agravitropic response at the young seedling stage and the lazy habit (prostrate growth of tillers) at the more advanced stage of growth. In this study, it was found that both agravitropism and lazy habit were controlled by the single recessivela gene. That is, F₂ segregants of Kamenoo×lazy-Kamenoo, which had an agravitropic response at their young seedling stage, showed a lazy habit of growth in the more advanced stage of vegetative growth. On the other hand, seedlings that showed normal gravitropic curvature at their early stage of growth had an upright growth in the mature stage.     

RNase and DNase activities in the alfalfa and lentil grown in iso-osmotic solutions of NaCl and mannitol

Nucleolytic activities from two plants of Leguminosae family were determined in order to consider if the nucleases of plants which belong to the same family or to the same species responded in similar ways to stress conditions during growth. Growth parameters of both plants were examined in parallel. In detail, seedlings from two plants, alfalfa (Medicago sativa L. cv. Luzerne Euver) and lentil (Lens culinaris cv. Thessalia), showed significant differences in response to iso-osmotic solutions of NaCl (100 mmol · L⁻¹ solution equivalent to conductivity 8.0 dS m⁻¹) and mannitol (190 mmol · kg⁻¹). Plant height and dry weight of mannitol/NaCl-treated seeds in both plants were lower in comparison to controls (water). Mannitol stress reduced height and dry weight in alfalfa seedlings more than did NaCl. By contrast, lentil seedling growth was inhibited more by NaCl stress than mannitol. In addition, DNase and RNase response to mannitol stress differed in each plant compared to the controls. Mannitol stress induced a sharp increase in DNase- and RNase-specific activity during the initial stages of alfalfa seedlings' growth, followed by a decrease during subsequent days; in lentil seedlings, these activities were inhibited throughout the entire growth period. NaCl stress inhibited the above activities in both plants. After native electrophoresis on gels polymerized in the presence of DNA/RNA, the overall band intensities confirmed the above quantitative results of alfalfa RNase and DNase activity. In addition, the active gel analysis revealed that the decrease of nucleolytic activities in mannitol-treated alfalfa seedlings was mainly due to the strong reduction of acid nucleases. This is the first report of different non-ionic osmotic response of type I plant nucleases during seedlings' growth. In vitro, the addition of up to 300 mmol/L mannitol did not affect acid and neutral nuclease activity in enzyme preparations extracted, purified, and separated from control and mannitol-treated alfalfa seedlings.Our results suggest that plant nucleases responded in a different way to osmotic stress and ionic stress conditions during seedlings' growth.     

Seed dispersal, plant recruitment and spatial distribution of Bactris acanthocarpa Martius (Arecaceae) in a remnant of Atlantic forest in northeast Brazil

Seed dispersal ecology of Bactris acanthocarpa Mart. (Arecaceae), an Atlantic forest understory palm, was investigated during two years as an attempt to test the following predictions: (i) seeds of Bactris are dispersed by mammals and large-gaped birds; (ii) Bactris benefits from seed dispersal in terms of seed predation avoidance, improvement of seed germination and seedling survival; and (iii) spatial distribution of adults is related to patterns of seed dispersal. The study was conducted at Dois Irmãos Reserve, a 387.4-ha reserve of Atlantic forest in northeastern Brazil (8º S–35º W). Black–rumped agoutis (Dasyprocta prymnolopha) and Guianan squirrels (Sciurus aestuans) were identified as the seed dispersers/predators, moving seeds short distances (< 4 m from parents) and at low rates (0.04-0.05 diaspore/palm/day). Pyrene burial prevented seed predation by vertebrates and reduced by half seed infestation by Scolytidae beetles. Only buried pyrenes germinated. Pyrene predation was not correlated with distance from conspecific adults. In contrast, early seedling mortality was higher near conspecific adults. Most adults (64%) had their nearest conspecific adult neighbour > 4 m away in contrast to 96% of seedlings that occurred concentrated within 4 m from adults (77% under the palm crowns). Here, we present evidence that spatial distribution of B. acanthocarpa is partly due to low rates of seed removal, short-distance seed dispersal by agoutis and squirrels, and early seedling mortality associated with presence of seedlings under palm crowns.     

Prediction of heavy-metal concentrations in mature plants by chemical analysis of seedlings

Five plant species were cultivated on a soil from the Neckar alluvial fan near Heidelberg (FRG) polluted by the emissions of a cement plant. Thallium, cadmium and lead concentrations in seedlings and mature plants were determined by atomic absorption analysis. AdditionallyBrassica napus L.napus was grown on soils containing 5 different concentrations of heavy metals, achieved by mixing two similar soils, from the same area but with different metal concentrations. Thallium and cadmium were shown to be taken up by roots whilst lead which was also absorbed, was deposited mainly on the plant surface. However during cultivation in the winter months, a remarkable deposit of lead via the roots was found. Thallium in the soil from a anthrorogen source was more available to plants than thallium of geological origin. During the lifetime of a plant concentrations of thallium and cadmium were always highest in the seedling. The decrease in metal concentration with maturity depended on the plant species and the element, but was not a function of the metal concentration in the soil.     

<Emphasis Type="Italic">Penicillium frequentans</Emphasis> isolated from<Emphasis Type="Italic"> Picea glehnii</Emphasis> seedling roots as a possible biological control agent against damping-off

Picea glehnii seedlings are affected by damping-off fungi in nurseries. The aims of this study were (1) to isolate fungi grown in the seedling rhizosphere in forest soil of P. glehnii, (2) to select fungi that produce antifungal compounds against Pythium vexans, and (3) to examine whether or not selected fungi can protect seedlings from P. vexans. Penicillium frequentans from Picea glehnii seedling roots produced antibiotic penicillic acid. Penicillic acid did not cause significant phytotoxicity to the seedlings. Penicillium frequentans increased the average percentage of surviving seedlings when inoculated together with Pythium vexans, but the increase was not significant. Vigorous mycelial growth of P. frequentans around seedling roots seems to be one of the mechanisms for protection, but the amount of penicillic acid was too low to show antifungal activity in the seedling rhizosphere.     

Herbivory and seedling performance in a fragmented temperate forest of Chile

Forest fragmentation alters plant-animal interactions, including herbivory. Relying manipulative experiments, we test if the reduction in insect herbivory associated with forest fragmentation translates into increased seedling growth and survival of three tree common species (Aristotelia chilensis, Cryptocarya alba and Persea lingue) in forest fragments and continuous forests in coastal Maulino forest, central Chile. Furthermore, we test if after protecting seedlings from herbivorous insects, plant performance is increased regardless of forest fragmentation. Nursery grown seedlings were transplanted into four forest fragments and a continuous forest during 2002. Insects, important herbivores in this forest, were excluded from half the seedlings by repeated applications of insecticides. Compared to continuous forests, in forest fragments, herbivory was reduced in all three species, seedling growth was greater in A. chilensis and C. alba but not in P. lingue, and survivorship was unaffected by herbivory or fragmentation in all three species. Protecting seedlings from insects reduced herbivory in the continuous forest to similar levels attained in the forest fragments. No change in herbivory results from by protecting seedlings in forest fragments. These results confirm that insects are important herbivores in the Maulino forest and also support the hypothesis that fragmentation can have strong indirect effects on plant communities as mediated through trophic interactions.     

香蕉园施用白花鬼针草的控草增效作用

[目的] 探索香蕉园施用白花鬼针草的可行性。[方法] 采用培养皿萌发试验生物测定法,以发芽率、发芽势、发芽指数、活力指数等化感效应指标评价白花鬼针草对蕉园4种优势杂草的化感作用,同时通过盆栽模拟试验探讨香蕉园施用白花鬼针草后杂草、香蕉和土壤三者的关系。[结果] 当白花鬼针草浸提液浓度为0.0125 g·mL^-1时,对马唐综合化感效应为促进作用,对短叶水蜈蚣、牛筋草、柔弱斑种草综合化感效应为抑制作用,白花鬼针草浸提液浓度为0.025~0.1 g·mL^-1,对4种受体杂草综合化感效应均为抑制作用;白花鬼针草处理具有降低种子发芽率,延缓种子发芽时间的作用,同时对杂草萌发后的鲜重有微弱促进作用,但这种促进作用较弱,综合化感效应表现为抑制作用。随着白花鬼针草茎、叶施用量的增加,控草增效作用不断提升,当施用量为400 g·株^-1时,控草增效作用最佳,对杂草综合株防效为78.99%,综合鲜重防效为70.60%,香蕉苗生物量增加19.79%,土壤有机碳、碱解氮、速效钾依次增加8.72%、10.36%、16.30%。[结论] 本研究初步探明了在香蕉园施用白花鬼针草具有防控蕉园优势杂草、提高土壤肥力和促进香蕉生长的效应。     

Storage reserve mobilization during low temperature germination and early seedling growth in Brassica napus

In western Canada, oilseed rape (Brassica napus L. var. oleifera cv. Westar) is seeded during the early months of spring, when ambient temperatures are well below the optimum. This can result in poor seedling emergence. The objectives of the present study were to determine which developmental stages are sensitive to low temperature and whether the effects are thermal or developmental in nature. Seed was germinated at 22, 10 and 6 °C. Fresh weight changes and seedling growth were assessed on the basis of equal accumulated heat units, and the mobilization of storage reserves was assessed by employing antibodies against isocitrate lyase (ICL; EC 4.1.3.1), oleosin and cruciferin. Additionally, de novo protein synthesis was determined by quantifying the incorporation of methionine via in vivo labelling. Low temperature resulted in poor germination and early seedling growth with phase II of germination being most sensitive. At 10 °C, there was a temporal delay in germination that did not affect the overall success of germination. This was a thermal effect as seed at the lower temperatures required the equivalent of 16–24 degree days before germination occurred. Also, seedling growth at 10 °C was lower in comparison to seedlings grown at 22 °C. Seed at 6 °C displayed slow and incomplete germination and poor seedling growth as a result of both thermal and developmental effects.     

过表达OsCatC基因水稻的获得及其耐盐性机理分析

过氧化氢酶C (Catalase C,CatC)作为重要的抗氧化酶,在水稻发育和胁迫响应方面起重要作用。为了探究CatC在盐胁迫响应中的功能及其作用机制,该研究构建了OsCatC过表达转基因水稻,并比较了其耐盐性和相关抗逆生理指标的变化。结果表明:(1)成功构建过量表达载体pCUbi1390-OsCatC-Flag,并经农杆菌介导的愈伤组织转化获得了30个独立转基因株系(T₀),Western blot鉴定T₁代幼苗共获得2个OsCatC过表达株系(OE-10、OE-18);qRT-PCR分析发现,OE-10和OE-18株系的OsCatC转录水平显著高于野生型(WT),证明OsCatC基因已成功过表达于转基因株系(OE-10、OE-18)中,且能正常翻译为融合蛋白CatC-Flag。(2)正常水培条件下,OE-10和OE-18与WT的水稻幼苗长势无明显差异,200 mmol·L^-1 NaCl处理7 d后再恢复水培10 d时,OE-10和OE-18幼苗的存活率为20%～25%,而WT幼苗绝大部分则干枯死亡,存活率仅为5%左...     

The allelopathic potential of alfalfa root medicagenic acid glycosides and their fate in soil environments

Summary The allelopathic effect of alfalfa (Medicago media Pers.) and red clover (Trifolium pratense L.) root saponins on winter wheat seedling growth and the fate of these chemicals in soil environments were studied. Seed germination, seedling and test fungus growth were suppressed by water and by alcohol extracts of alfalfa roots, and by crude saponins of alfalfa roots, indicating that medicagenic acid glycosides are the inhibitor. Powdered alfalfa roots inhibited wheat seedling growth when added to sand. At concentrations as low as 0.25% (w/w) the root system was completely destroyed whereas seedling shoots suffered little damage. Red clover roots caused some wheat growth inhibition when incorporated to sand, but their effect was much lower than in the alfalfa root treatment. Soil textures had a significant influence on the inhibitory effect of alfalfa roots. The inhibition of seedling growth was more pronounced on light than on heavy soils. This was attribted to the higher sorption of inhibitors by heavy soils. Incubation of alfalfa roots mixed into loose sand, coarse sand, loamy sand and clay loam for a period of 0–8 days resulted in decreased toxicity to bothT. viride and wheat seedlings. This decrease occurred more quickly in heavier soils than in loose sand, due to the hydrolysis of glycosides by soil microorganisms. Soil microbes were capable of detoxifying medicagenic acid glycosides by partial hydrolysis of sugar chain to aglycone. These findings illustrate the importance of medicagenic acid glycosides as an inhibitor of wheat seedling growth, and of their fate in different soil environments.     

3种菊科入侵植物不同生长时期的土壤酶活性和养分变化

【目的】黄顶菊、三叶鬼针草和豚草是我国危害较严重的3种菊科入侵植物。了解3种菊科植物生长过程中土壤养分和酶活性的变化,可以为研究其入侵机制提供依据。【方法】在中国农业科学院植物保护研究所廊坊中试基地开展同质园实验,比较分析了黄顶菊、三叶鬼针草和豚草3种菊科入侵植物在幼苗期、旺盛生长期和生殖生长期根际土壤养分和酶活性的变化。【结果】3种外来菊科植物的生长时期对土壤养分和酶活性存在显著影响。3种外来植物入侵域的土壤速效磷、硝态氮含量以及磷酸酶活性随生长时期变化都表现出先升高后降低的趋势,均在旺盛生长期达到最大值。入侵域土壤速效磷和硝态氮含量的变化趋势与狗尾草明显不同。3种外来植物入侵降低了土壤速效钾含量,提高了土壤脲酶、磷酸酶活性。【结论】3种外来菊科植物的入侵改变了土壤养分和酶活性,创造出更有利于自身生长和繁殖的条件,以利于其进一步扩张。     

林隙对香樟和枫香幼苗早期生长阶段功能性状的影响

探究林隙对不同需光性树种早期生长特征和功能性状的影响,对揭示林隙微生境影响次生林内幼苗更新机制具有重要意义。以亚热带次生林中耐荫常绿树种香樟和阳性落叶树种枫香幼苗为试验对象,研究大林隙(D/H介于1.5—2.0)、中林隙(D/H介于1.0—1.5)和小林隙(D/H介于0.5—1.0)对不同需光树种幼苗早期(1—3年生)生长特征和功能性状的影响。结果表明：(1)林隙大小对两种幼苗的生长均有显著影响。其中,中林隙可显著促进香樟2—3年生幼苗的生长,大林隙对枫香1—3年生幼苗的生长均具有显著促进作用。(2)对林隙环境因子与幼苗功能性状的关系进行冗余分析表明,香樟幼苗功能性状的变化与林隙土壤有机质含量、水解性氮含量、酸碱度和有效磷含量密切相关,而枫香幼苗功能性状则主要受林隙土壤酸碱度、有机质含量、水解性氮含量、土壤含水率、冠层透光率和土壤有效磷含量的影响。(3)维持较高的根重比、细根比根长、叶碳氮比和叶碳磷比是幼苗应对林隙环境影响的重要生理生态调节机制。     

Chlorophyll content and leaf elongation rate in wheat seedlings as a measure of manganese tolerance

After aluminum toxicity, manganese (Mn) toxicity is probably the second most important growth limiting factor in acid soils. The purpose of this study was to determine the feasibility of using chlorophyll content and leaf elongation rate (LER) for regrowth of Mn stressed seedlings as a rapid seedling based screening bioassay for Mn tolerance in segregating populations of wheat (Triticum aestivum L.). In one experiment, chlorophyll was determined for the cultivars Norquay (Mn-tolerant) and Columbus (Mn-sensitive) subjected to twelve Mn levels (2 to 2000 μM) in nutrient solutions. As Mn concentration increased, chlorophyll ‘a’ and ‘b’ contents of the Mn-tolerant cultivar decreased up to 9%, while in the Mn-sensitive cultivar it was reduced by as much as 43%. The chlorophyll ‘a/b’ ratio did not differ among Mn concentrations for either cultivar. In a second experiment, chlorophyll content and LER for regrowth of Mn stressed seedlings (1000 μM) was determined for Columbus and Katepwa (Mn-sensitive), Oslo (Mn-intermediate), and Norquay and Laura (Mn-tolerant). Manganese tolerance as assayed by chlorophyll ‘a’ and ‘b’ and LER was significantly correlated with Mn tolerance as assayed by the relative root weight methodology (RRW). Thus, chlorophyll content of Mn-stressed seedlings and LER of seedling regrowth appear to be suitable techniques for screening unreplicated selections of segregating populations for tolerance to Mn.