山羊骨髓MSCs体外向成骨细胞诱导和分化

以关中奶山羊骨髓间充质干细胞（MSCs）为对象，研究了体外诱导山羊MSCs向成骨细胞的分化。结果表明，山羊MSCs在地塞米松、β-磷酸甘油和抗坏血酸的作用下，从诱导的第4~5天开始，即可见有些MSCs体积逐渐增大，由梭形或多角形转变为立方形或卵圆形；随着诱导时间延长，立方形或卵圆形细胞不断增多，到第15天时，有较多的立方形细胞，且这种变化经历了从立方形向卵圆形转变的过程；第25天，可见卵圆形和方形细胞堆积，诱导分化率为90.2% ± 2.97%。说明其被诱导分化为成骨细胞表型，该类细胞呈茜素红染色和碱性磷酸酶(ALP)染色阳性。从而证明了山羊骨髓MSCs具有体外分化为成骨细胞的潜能，从而为利用该类细胞作为实验模型研究骨骼缺损修复等提供了科学依据。     

猪羊水干细胞的分离培养及向脂肪细胞诱导分化

羊水干细胞是一种存在于胎儿羊水中Oct4阳性的多能性干细胞.该细胞具有增殖能力强、分化潜能高和免疫原性低等特点,在生物学和医学研究上具有广泛的应用前景.本实验从不同胎龄的猪胎儿标本中分离猪羊水干细胞,并通过诱导其向脂肪细胞分化来检测其多向分化潜能.结果表明,从胎龄为30～70 d的猪胎儿羊水中分离出的羊水干细胞贴壁率高,增殖能力强,活性好,并建立了两株猪羊水干细胞系.经流式细胞仪和RT-PCR检测,猪羊水干细胞表达CD117、CD44、CD166和CD90表面抗原,不表达CD45、CD54和CD34;稳定表达Oct4、Nanog、C-myc和HLA-abc干细胞特异标志基因;转染报告基因载体pOct4-EGFP和pNanog-EGFP后表达绿色荧光;此外,猪羊水干细胞还能够在体外诱导分化为脂肪细胞.研究结果为大动物羊水干细胞的研究提供了可借鉴的材料.     

小鼠生殖细胞特异基因Oct4启动子片段克隆及报告载体构建

本研究通过PCR方法从小鼠(Mus musculus)基因组中扩增出Oct4启动子的CR4区和CR1区,将其克隆到真核报告载体pEGFP-1,构建了携带Dct4启动子片段的小鼠生殖细胞特异性报告载体CR1,4-pEGFP-1.用该载体转染多种细胞,在荧光显微镜下观察绿色荧光蛋白(EGFP)在细胞中的表达情况,同时以半定量RT-PCR检测转染细胞中Oct4及其它生殖细胞特异性基因的转录情况.结果显示,GFP在小鼠胚胎干细胞、小鼠胚胎成纤维细胞(MEF)和C2C12细胞中均不表达,而在P19细胞和睾丸细胞中表达.RT-PCR结果显示,Oct4在P19、睾丸细胞和生殖干细胞中特异性转录.这表明所构建的绿色荧光蛋白报告载体只在生殖细胞和多能生殖干细胞中内表达,该质粒可作为示踪生殖细胞的工具,同时也可以作为生殖干细胞多能性状态的一个监测标记.     

猪多能干细胞建系的培养条件及其分子调控网络研究

多能干细胞(pluripotent stem cells,PSCs)作为一类重要的细胞资源,成为目前国内外的研究热点,获得这类细胞在当今医学研究和生物技术的发展及应用中具有重要的意义。猪(Sus scrofa)由于其生理结构和代谢特征与人(Homo sapiens)较为相似,成为动物多能干细胞建系的重点研究对象。然而目前,猪多能干细胞建系工作仍存在很多难题。本文将从猪多能干细胞建系研究的进展和存在的问题入手,对照小鼠(Mus musculus)和人相关研究的成功经验,总结目前已经报道的猪多能干细胞的培养条件、调控因子、信号通路和基因表达特征,以期为建立猪胚胎干细胞和诱导多能干细胞系提供参考。     

人源羊水干细胞的培养、鉴定及影响因素分析

取人羊水标本进行羊水干细胞原代,分离培养,并探讨人羊水标本性状等因素对原代分离培养的影响.结果表明,分离的羊水干细胞表达Oct-4等胚胎干细胞和CD29等成体干细胞的特异基因,细胞生长迅速,增殖能力较强,并可分化为Nestin阳性的神经细胞和α-actia阳性的心肌细胞.同时,在羊水干细胞原代培养时,羊水标本性状在一定程度上影响细胞的贴壁数量及贴壁时间.羊水标本细胞的贴壁时间孕中期(4.7±0.6)d与孕晚期(6±0.5)d有明显差异(P<0.05),孕晚期贴壁时间较长.羊水中血液污染程度对贴壁时间有明显影响,重度污染组(10.8±0.3)d与无污染组(6±0.5)d、轻度污染组(6.3±0.6)d贴壁时间差异均显著(P<0.05).羊水体积对原代细胞的贴壁数量和细胞贴壁时间有一定影响,羊水体积增大贴壁时间延长,但差异不显著,而贴壁细胞数量随羊水体积增加而增加,各组之间差异显著(P<0.05).实验系统地检测了羊水干细胞原代培养的影响因素,为羊水干细胞研究提供了可以借鉴的资料,同时分离得到的羊水干细胞,具有分化为神经细胞和心肌细胞的潜能,有望作为种子细胞进行临床应用.     

Dvl1及其突变体腺病毒载体的构建与在MSCs中表达鉴定

Dvl1及其结构缺失突变体ΔDIX（dsh and axin）和ΔDEP（dsh,Egl-10 and pleckstrin）腺病毒载体的构建并验证其在MSCs（mesenchymal stem cells）中的表达情况。将目的基因定向克隆至pAdTrack-CMV穿梭载体上,并经PmeⅠ线性化后在BJ5183细菌中与pAdEasy-1骨架质粒同源重组,获得重组腺病毒载体,在QBI-293A细胞中包装及扩增,实时荧光定量PCR和Western bolt验证Dvl1在MSCs中的表达情况。经PCR、PacⅠ单酶切鉴定及测序分析,成功构建Ad-Dvl1、Ad-ΔDIX和Ad-ΔDEP腺病毒载体,目的基因序列与GenBank报道一致,并选出150 MOI为最适感染MSCs的感染复数,成功构建了Ad-Dvl1、Ad-ΔDIX和Ad-ΔDEP腺病毒载体,并获得高滴度的病毒子,qPCR和Western blot证实Dvl1在MSCs中高表达并增加了β-catenin在细胞中的累积,为进一步研究Dvl1在MSCs迁移过程的作用奠定了基础。     

国产丝裂霉素处理的STO细胞作为饲养层培养牛胚胎干细胞

寻找一种适合牛胚胎干细胞生长的饲养层对成功培育牛胚胎干细胞有重要意义.本研究采用不同浓度国产丝裂霉素处理的SIM小鼠(Mus musculus)成纤维细胞耐硫代鸟嘌呤和耐鸟苯苷亚系(STO细胞)作为饲养层培养牛胚胎干细胞,为牛胚胎干细胞培养体系的建立提供实验依据.用10、15、20、30和40 μg/mL国产丝裂霉素分别处理1.5、3.0和4.0 h的STO细胞;形态学观察不同代次STO细胞生长状态,并对生长在以STO细胞为饲养层的牛胚胎干细胞进行碱性磷酸酶染色(AKP)、阶段特异性胚胎细胞表面抗原-4(OCT-4)和阶段特异性胚胎抗原-1(SSEA-1)免疫组化检测、体外分化形成拟胚体等实验.结果显示,当丝裂霉素的浓度为15 μg/mL处理3.0 h可有效的抑制细胞分裂;STO细胞在5～10代作为饲养层细胞形态最好且获得的牛胚胎干细胞AKP染色及OCT-4和SSEA-1抗原表达均成阳性,分别在体外分化培养形成了拟胚体.结果证明使用国产丝裂霉素能有效地处理STO细胞且在以STO细胞作为饲养层培养的牛胚胎干细胞能保持未分化状态,可以作为常规饲养层培养牛胚胎干细胞.     

家畜多能干细胞的研究现状与应用

多能干细胞(pluripotent stem cells,PSCs)包括从胚胎内细胞团中分离出的胚胎干细胞(embryonic stem cells,ESCs),外胚层干细胞(epiblast stem cells,EpiSCs),从原始生殖细胞(primordial germ cells,PGCs)分离出的胚胎生殖细胞(embryonic germ cells,EGCs)和诱导多能干细胞(induced pluripotent stem cells,iPSCs).这些细胞都具有向各种类型细胞发育的潜能,因此,PSCs为再生医学的发展,人类基因疾病的动物模型的建立,异种器官移植和高质量抗病新品种的培育提供了有效的途径.迄今为止,PSCs在小鼠(Mus musculus)模型和人类物种已经做了大量研究,同时,家畜的ESCs和iPSCs也取得了有效的进展,表明其不仅可以应用于特定家畜优良品性的稳定遗传和优育,而且可以以家畜为载体生产人类需要的化学药品和抗体,甚至应用在人类疾病的临床前应用中.然而,家畜的ESCs相对来说是难分离和富集的,体外培养建立稳定的ESCs系具有更大的难度和挑战.近年来,得益于对家畜ESCs和相关iPSCs相关的持续研究,已初步揭示了家畜ESCs的独特生物学特性.PSCs已经成为生命科学和高科技农业和生物学一个创新的研究领域.在本文中,本文综述了家畜PSCs的发展历程与应用前景.     

L-Wnt3a细胞用于制备胚胎干细胞饲养层的研究

小鼠胚胎成纤维细胞(mouse embryonic fibroblasts,MEFs)作为胚胎干细胞培养的饲养层细胞,受到细胞代次等因素限制,需要长期反复制备,消耗大量的实验动物。寻找可替代小鼠胚胎成纤维细胞支持干细胞生长增殖的稳定细胞系,对于降低干细胞的培养成本和减少工作量具有重要意义。L-Wnt3a细胞是一个能够稳定向细胞外分泌Wnt3A蛋白的永生细胞系,目前尚不清楚其是否支持小鼠胚胎干细胞的培养。本研究探讨L-Wnt3a作为饲养层细胞对于小鼠(Mus musculus)胚胎干细胞系的建立以及该细胞条件培养液对无饲养层培养小鼠胚胎干细胞的影响。实验结果表明,丝裂霉素C浓度为10μg/mL,作用4 h时,可显著抑制L-Wnt3a细胞的体外生长而不影响细胞的活力。用该细胞制备饲养层,原代分离、培养小鼠胚胎干细胞,成功建立小鼠胚胎干细胞系。利用L-Wnt3a细胞制备条件培养液进行胚胎干细胞的培养,结果显示,L-Wnt3a条件培养液可支持小鼠胚胎干细胞在无饲养层条件下自我增殖,该条件下培养的胚胎干细胞表达碱性磷酸酶、Oct4、Sox2、Nanog、E-cadherin(E-CAD)和SSEA-1多能性因子,并能够在体内、外分化为3个胚层,形成嵌合体后代。L-Wnt3a细胞作为饲养层细胞,能够成功分离获得小鼠胚胎干细胞,可以避免小鼠胚胎成纤维细胞作为饲养层细胞存在的代数限制、制备繁琐等问题;并且LWnt3a条件培养液可以在无饲养层条件下维持小鼠胚胎干细胞的自我更新,能够在一定程度上有效避免饲养层细胞对胚胎干细胞造成的不利影响。     

细胞非对称分裂与姐妹染色体非随机分离研究进展

细胞非对称分裂与染色体非随机分离一直是生命科学研究领域的热点,细胞非对称分裂包含形态学非对称分裂与功能性非对称分裂,对于生命世代延续与维持个体组织功能完整意义重大.近年来,在生殖细胞与干细胞研究领域,对于细胞非对称分裂现象的研究取得了一些新的进展,这些进展有助于理解和认识细胞非对称分裂的形成及其调控机制.哺乳动物卵母细胞成熟过程中非对称分裂(形成较大体积的卵子和较小体积的极体)是形态学非均等分裂的代表;而损伤所诱发的干细胞的增殖则是细胞功能性非对称分裂的代表,这一过程所产生的两个后代细胞尽管形态近似,但是具有不同的命运与功能,一个留在原来微环境中继续维持干细胞特性,另外一个则分化成为功能性细胞,替代坏损细胞的功能.细胞在形态学与功能性的非均等分裂,其根源都是细胞核物质的选择性分离.果蝇(Drosophilamelanogaster)的精巢组织可在体外进行活体显微观察,能够对精原干细胞的细胞非对称分裂与姐妹染色体分离的实时观察研究,近年来细胞非对称分裂与姐妹染色体分离的研究结果主要来自果蝇.本文结合近年来关于果蝇精原干细胞的姐妹染色体的非对称分离的研究,对细胞非对称分离的假说、研究手段及其可能的调控机制等进行回顾总结,希望能为该领域的未来发展提供一些思路与借鉴.     

“绿色革命”以来大气CO2浓度升高对C3作物营养品质的影响

自20世纪60年代"绿色革命"以来,育种技术和农耕技术的发展促进了农作物产量的大幅提升,然而作物的营养品质出现下降趋势。在相似的遗传背景下,大气CO_2浓度升高会使单位体积农作物产品的营养元素含量下降,因此"绿色革命"至今,农作物产品的营养元素下降可能受大气CO_2浓度升高影响。通过植物生长箱模拟"绿色革命"初期和目前的大气CO_2浓度水平(310μmol/mol和400μmol/mol),针对主要C_3作物水稻、小麦和大豆,研究"绿色革命"以来大气CO_2浓度升高对其籽粒的C、N、Fe、Zn元素含量的影响,结果表明:CO_2浓度升高对3种作物籽粒的C元素含量几乎没有影响,变化幅度在±1.5%之间;籽粒的N、Fe、Zn元素含量普遍呈现下降趋势,但均未达到显著水平。     

Potential of wild species for genetic enhancement of some semi-arid food crops

Discovery and incorporation of genes from wild species provide means to sustain crop improvement, particularly when levels of resistance in the cultigens are low and virulent strains of pests and pathogens overcome the host plant resistance. The extent of utilization and the potential of the wild genepool for genetic enhancement were reviewed in five important food crops viz. sorghum, pearl millet, chickpea, pigeonpea and groundnut grown in the semi-arid tropics. Introgression from compatible wild germplasm in the primary gene pool resulted in transfer of new cytoplasmic male sterility systems in pearl millet and pigeonpea, development of high protein, cleistogamous flower and dwarf pigeonpea lines and foliar disease resistant groundnut cultivars. Utilization of wild species in secondary and tertiary gene pools has been generally limited due to sterility, restricted recombination or cross incompatibility. Nevertheless, these species are extremely important as they contain high levels of resistance to several important biotic and abiotic stresses. Several of them, like those belonging to the Parasorghum section and the rhizomatous Arachis species are sources of multiple resistances and hold great promise to sustain crop productivity.     

Faculty positions:Center for Agricultural Resources Research,Chinese Academy of Sciences

正The Center for Agricultural Resources Research(CARR),the Institute of Genetics and Developmental Biology(IGDB),Chinese Academy of Sciences,invites applicants for several research group leader positions.CARR is one of the research organizations in Chinese Academy of Sciences(CAS).We seek nominations and applications     

Soil-profile distribution of primary and secondary plant-available nutrients under conventional and no tillage

Nutrient distributions under no tillage (NT) compared with conventional disk-and-bed tillage (CT) management in the warm, humid region of the southeastern USA need to be assessed so that future placement, quantity, and type of fertilizers can be altered, if necessary, to efficiently match crop demands. We determined soil-profile distributions of pH, N, P, S, K, Ca, Mg, Na, Zn, Fe, Mn, and Cu to a depth of 0.9 m at the end of 8.5 years of continuous CT and NT management on a Weswood silty clay loam (fine, mixed, thermic Fluventic Ustochrept) in southcentral Texas. Most dramatic changes occurred within the 0–0.05 m depth, where soil under NT had lower pH, Fe, and Cu than under CT, but greater P, K, Zn, and Mn. Greater P and K under NT than under CT also occurred below the till-zone (0.15–0.3 m). At a depth of 0–0.3 m, soil under NT contained greater amounts of extractable P, K, Zn, Fe, Mn, and Cu than under CT. Nitrogen fertilization had little effect on nutrient distributions, except resulting in greater extractable K at 0–0.05 m and greater nitrate at 0–0.15 m. Few changes in soil-profile distributions were observed for extractable S, Ca, Mg, and Na. Long-term continuous use of NT on this fine-textured, high-fertility (except for N) soil had no apparent adverse effects on nutrient distributions relative to CT, but enhanced conservation and availability of P, K, Zn, Fe, Mn, and Cu near the soil surface where crop roots proliferate.     

Assessing the effectiveness of conservation measures: Resolving the “wicked” problem of the Steller sea lion

“Wicked” problems are those that are complex and that change when solutions are applied. Many conflicts in conservation fall into this category. The study approached the problem of how to constrain the apparent wickedness of a problem in the conservation management of a species by using simple empirical indicators to carry out iterative assessment of the risk to a population and to document how this risk evolves in relation to the addition of new data and the implementation of management actions. Effects of high levels of uncertainty within data and also concerning population structure were examined through stochastic simulation and by exploration of scenarios. Historical trends in the example used, the Steller sea lion, showed rapid declines in abundance in some regions during the 1980s. The current total population is 130,000-150,000 Steller sea lions through Alaska and British Columbia and this number has been stable since about 1990 in spite of regional differences in population dynamics. Regional differences in the sequence of changes in the number of pups and non-pups, suggested that an internal re-distribution of juveniles could have happened between 1980 and 1990. Current productivity also appears close to the long-term mean. Stochastic population projection using various scenarios showed that, based upon this history, the risk of extinction for the population has declined and is below reasonable thresholds for considering the population to be endangered. The trends in risk suggest that management actions taken since 1990 have probably been effective. Consequently, the conservation management objectives for the Steller sea lion are probably being met. The approach provides a mechanism, based upon experience and scenario analysis, for exploring future policy options and may help to constrain the debate amongst stakeholders about the cost-benefit trade-offs associated with different options.     

Collection and preliminary evaluation of native turfgrass accessions in Italy

Despite a raising interest on turfs in Italy, all theavailable varieties of this kind in the Country are of foreign origin, and areoften poorly adapted to the prevailing climatic conditions. This prompted tobegin a collection activity of indigenous turfgrass species, with the ultimategoal of identifying promising materials for future breeding based on localgenetic resources. The collection was carried out in three areas of Italy, viz.the northern Po Plain, the coastal region of Liguria, and the island of Sardiniathat are characterised, respectively, by subcontinental, warm temperate, andtypical Mediterranean climate. Altogether, 141 sites were visited, yielding 226accessions belonging to eight species of potential interest for turfs:Poa pratensis, Poa trivialis,Festuca rubra, Festuca arundinacea,Lolium perenne, Agrostis stolonifera,Agrostis tenuis, and Cynodon dactylon,this last being a warm-season grass. Poa pratensis andCynodon dactylon were mostly collected in northern Italyand Sardinia, respectively, whereas Festuca arundinacea andLolium perenne were rather ubiquitous. The collection sitesranged from 0 to 1040 m asl, but sites over 750 m wereonly visited in the inner part of Sardinia. All the accessions, collected aswhole plants, were transplanted at Lodi, northern Italy, where they are beingevaluated. Their preliminary evaluation for traits of importance for turf use,such as sward colour and overall quality, highlighted the great variation andthe occurrence of interesting accessions in all species. Other characters wererecorded, bearing specific importance in individual species, and in all casespromising accessions were identified. The germplasm of Festucarubra, Festuca arundinacea, and Loliumperenne proved highly infected by endophytic (symbiotic) fungi of thegenus Neotyphodium.     

Enhanced establishment and growth of giant cardon cactus in an eroded field in the Sonoran Desert using native legume trees as nurse plants aided by plant growth-promoting microorganisms and compost

To evaluate the feasibility of long-term desert reforestation technology of mixed vegetation, cardon cactus (Pachycereus pringlei) seedlings from indoor and outdoor nurseries were planted in the field adjacent to one seedling of potential legume nurse trees: mesquite amargo (Prosopis articulata), yellow palo verde (Parkinsonia microphylla), and blue palo verde (Parkinsonia florida). Some of the planting holes were also supplemented with common dairy compost. Additionally, the combinations of legume tree–cactus were inoculated with either a consortium of desert arbuscular mycorrhizal (AM) fungi, plant growth promoting bacteria (PGPB; the diazotroph Azospirillum brasilense Cd, and the phosphate solubilizer Paenibacillus sp.), or a mixture of all. The field experiments were evaluated periodically during 30 months for survival and growth. Cardons reared in an outdoor screen house survived better in the field than those reared in a controlled growth chamber and hardened later outdoors. Association with any legume nurse tree increased survival and enhanced growth of untreated cardons. For cardons growing alone, application of either compost, AM fungi, and all the treatments combined increased survival. For these plants, no treatment affected plant growth during the first 3 months after transplanting. Later, all treatments, except for AM fungi, enhanced plant growth. However, only 2 years after transplanting the enhanced growth effect of AM fungi was also significant. In the presence of the legume nurse trees, transient positive effects on cardon growth were recorded. General evaluation after 30 months of cultivation showed that the treatments positively affected cardon growth when growing alone or in combination only with mesquite amargo but not with the other two legume trees. This study proposes that young legume trees have the capacity to enhance survival and growth of cardon cactus, depending on the legume cactus combination. Additional treatments such as compost or PGPB can either amplify the effect or else attenuate it.     

Karyotypic and genome size variation in Cajanus cajan (L.) Millsp. (pigeonpea) and some wild relatives

Detailed karyotypes and 4C DNA amounts have been studied in five cultivars of Cajanus cajan and 20 species belonging to Cajanus, Rhynchosia, Dunbaria, Flemingia and Paracalyx. C. cajan shows intraspecific variability and its karyotype is most similar to that of C. cajanifolius (sect. Cajanus) in the morphology and number of satellite chromosomes and the lack of any correlation between chromosome size within the complement and asymmetry. Karyotypes of C. lineatus and C. sericeus belonging to sect. Atylia are similar with respect to maximum r-index and the ratio of longest and shortest chromosomes in their respective complements. C. acutifolius (sect. Frutocosa) is distinct in having a very low ratio (1.44) between the longest and the shortest chromosomes, while in C. albicans, C. goensis, C. scarabaeoides (sect. Cantharospermum) there is no chromosome pair with r-index >2.0. C. mollis and C. volubilis (sect. Volubilis) show similarity with regard to ratio between the longest and the shortest chromosomes and a distinct chromosome pair with an arm ratio of 2.7 and C. platycarpus is distinct (sect. Rhynchosoides) in having the smallest ratio (1.36) between the longest and the shortest chromosomes. 4C DNA amounts in Cajanus and Rhynchosia vary between 3.28 pg to 11.69 pg and 4.92 pg to 11.13 pg respectively, while in Dunbaria, Flemingia and Paracalyx these vary between 4.70 and 7:06 pg. In the genus Cajanus karyotypic features and 4C DNA amounts agree with the sectional classification and bring out a close relationship between C. cajan and C. cajanifolius. This is supported by studies based on seed protein patterns, isozyme analysis, trypsin and chymotrypsin inhibitor patterns, RFLP and RAPD data and crossability relationships. A clear relation between DNA amount and abnormalities effecting the extent of pairing and recombination in interspecific hybrids shows the importance of this study in developing future breeding programme involving C. cajan and its wild relatives that are of potential value.     

Effect of some fungicide seed treatments on the survival of Bradyrhizobium japonicum and on the nodulation and yield of soybean [Glycine max. (L) Merr.]

Several commercial fungicide seed treatments were evaluated for their possible effect on the survival of Bradyrhizobium japonicum on seeds and on the nodulation and yield of soybeans in a greenhouse and a field experiment. quinolate Pro (carbendazim and oxine copper), Vitavax 200FF (carboxin and thiram), and Monceren (pencycuron) had a small effect or no effect on the survival of B. japonicum and on the nodulation and yield of soybeans. They can thus be considered compatible with soybean seed inoculation. Germipro UFB (carbendazim and iprodione), Apron 35J (metalaxyl), and Tachigaren (hymexazol) decreased B. japonicum survival and the nodulation and yield of soybeans and thus cannot be considered compatible with soybean seed inoculation.     

Leaf position and genotype differences for mineral element concentrations in sorghum grown on tropical acid soil 1

Mineral element deficiencies and toxicities are common problems associated with sorghum [Sorghum bicolor (L.) Moench] production on acid soils. To better understand some of the mineral element problems and the analysis of plant tissue of sorghum plants grown on acid soils, four sorghum genotypes were grown on an acid Oxisol at Carimagua, Colombia limed with dolomite at 2 and 6 Mg ha^‐1.

Samples for mineral element analyses were obtained from leaves at different positions on the four genotypes. Concentrations of P and Mg were highest in the flag leaf (Leaf No. 1) and decreased as the position on the plant declined from the top of the plant for plants grown at 2 Mg lime ha^‐1. Similar decreases in P, Mg, K, and Zn concentrations occurred in plants grown with 6 Mg lime ha^‐1. Concentrations of Ca, S, Si, Mn, Fe, Cu, and Al increased as leaf position declined from the flag leaf for plants grown at 2 and 6 Mg lime ha^‐1. The higher lime supply enhanced Ca and reduced Mn and Fe concentrations in leaves. Differences in mineral element concentrations for the four genotypes used were fairly extensive. The elements to show the greatest range among genotypes were Al and Si and the elements to show the least range among genotypes were P, K, and S. Care should be used in collecting leaf samples for plant analysis and genotypic differences for accumulation of mineral elements should be considered in interpretation of results.