986 resultados para photoperiod-sensitive genie male-sterile mutant
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近年来作物杂种优势利用的研究取得了很大的进展,杂交种的应用带来了巨大的经济效益。作物杂种的生产通常借助于雄性不育系。但是某些植物雄性不育形成的分子机理尚未搞清,不育基因的结构与功能以及不育基因在表达过程中一系列的基因与蛋白质的相互作用有待揭晓。 显性雄性核不育小麦(太谷核不育小麦)是我国特有的显性雄性核不育无花粉型材料,其不育性是由显性单基因Ms2控制的。本论文以显性雄性核不育小麦不育株和可育株近基因系为材料,应用单向电泳(SDS-PAGE)和双向电泳(IEF/SDS-PAGE)方法,分析了不育株和可育株不同器官(种子、旗叶、幼穗及花药)的蛋白质组成。通过研究发现两者之间在蛋质组成上存在一些异同点。在胚乳和旗叶中,未发现不育株和可育株两者之间的蛋白质组成存在明显的差异。在外于减数分裂时期的幼穗和花药中,不育株和可育株之间蛋白质组成上有明显差异。与可育株相比,不育株缺少分子量分别为15.8kD、17kD、17.8kD、38kD、81.2kD的5个碱性蛋白质。另外,还发现不育株增加了一个16.2kD的低分子量酸性蛋白质。在某些特定分子量的蛋白质中,虽然两者都存在该蛋白质,但是在含量上有着明显的不同,可育株含量比不育株高。这些蛋白质组成的变化成为不育株的重要特征。本文首次报告了显性雄性核不育小麦不育株与近等基因系可育株不同器官中蛋白质组成的区别。本研究结果在蛋白质水平上证实了花器官是雄性败育基因表达的主要器官,Ms的表达具有较强的时间性和空间性。 本论文还对双向电泳方法进行了一些探讨。
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植物通过异戊二烯代谢途径合成多种具有生物活性和功能的三萜及甾醇类化合物,它们在调节植物生长发育、维持膜的完整和功能、抵抗病原微生物侵染中发挥着重要的作用。2,3-氧化鲨烯为三萜和甾醇合成途径的分枝点,参与这一关键步骤的酶被通称为2,3-氧化鲨烯环化酶(OSCs)。本研究系统分了水稻基因组中全部11个OSC基因序列,发现其中四个可能为假基因。亚种间非同义替换率Ka和同义替换率Ks的比值(Ka/Ks)以及进化树的分析表明OsOSC8是单子叶植物特有的功能保守基因,而OsOSC9在水稻两个亚种间发生了功能快速进化,这种快速进化的基因往往参与植物和病原菌相互作用的代谢途径。 根据基因结构、表达谱以及与其它植物已知功能的OSC酶氨基酸序列的比对推测OsOSC3可能具有环阿屯醇合成酶的功能,参与植物甾醇的合成,而OsOSC7、OsOSC10和OsOSC11可能具有β-香树素合成酶的功能,其余OSCs可能参与合成其它三萜化合物。为了进一步分析和验证OSCs酶的功能,将水稻7个OSC基因的开放阅读框(ORF)构建到酵母表达载体并在pichia酵母中表达,发现仅有OsOSC9和OsOSC12能够将酵母内源的2,3-氧化鲨烯分别环化为四环三萜化合物Parkeol和植物中稀有的五环三萜化合物Isoarborinol,目前还未在其它植物中发现参与这两种三萜化合物的基因。另外,水稻所有的OSC基因均不能互补酵母羊毛甾醇缺陷型菌株,表明水稻OSCs不具有合成羊毛甾醇的功能。 RNAi沉默以及启动子融合GUS的表达实验发现OsOSC8可能参与花粉的发育,该基因的下调影响水稻的育性,暗示水稻中存在一个可能与雄性不育有关的三萜代谢途径。水稻其它OSC基因RNAi植株可能在逆境环境和病原菌侵染下才会显现出表型。
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The present study was conducted to assess the potential toxicity of the effluent from a large sewage treatment plant (GBD-STP) in Beijing. Japanese medakas (Oryzias latipes) at reproduction active period were exposed to a serial of graded concentrations of the effluent or 100 ng l(-1) of 17-alpha-ethinylestradiol (EE2, positive control). Growth, gonadosomatic index (GSI), hepatosomatic index (HSI), reproductive success, induction potency of vitellogenin (VTG) in male fish and that of 7-ethoxyresorufin-o-deethylase activity (EROD) in male fish liver were used as test endpoints. The growth suppression of fish was observed in a dose-dependent manner, resulting in significant differences in both body length and body weight of medaka above 5% effluent. This effluent can inhibit the growth of gonad of medakas and are more sensitive to male than to female. At exposure concentration of 40% and higher, there was an unexpected decrease of HSI values, which may be resulted from sub-lethal toxicity of effluent to fish liver. VTG of plasma in males were induced in all exposure concentration levels, but not in a dose-dependent manner. The concentration of 5% effluent would be the lowest observed adverse effect level (LOAEL) affecting reproductive success when examining fertile individuals, fecundity and fertilization rate. The overt CYP1A response and higher reproductive toxicity may be indicative of low process efficiency of this STP. (c) 2004 Elsevier Ltd. All rights reserved.
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水稻是重要的粮食作物,其产量的增加和品质的改良都是关系国计民生的大事。就我国现阶段的国情而言,水稻产量在现有水平上稳步提升仍是未来十几年甚至几十年农业生产最重要的目标之一。尽管根据“超级杂交水稻育种”的战略设想和水稻育种实践,通过不断地改进育种技术可望在更高的产量水平上进行水稻杂种优势利用,在稻属植物内还具有很大的产量潜力可以挖掘。然而,仅仅从现有的种质基础出发,要更大幅度提高水稻单产,实现“超级杂交稻”的目标也存在一些困难:现有的推广品种是二倍体,尽管种类众多,但是其基因组的来源相对单一;同时,水稻基因组DNA含量也是作物中最少的,基因组内寻求开发潜力有一定困难;水稻作为C3植物,光合利用效率不高也是制约水稻产量提高的因素之一。因此,寻求常规手段以外的技术突破或者方法创新,是实现“超级杂交稻”的目标的迫切需求。本研究利用秋水仙素能抑制细胞分裂中纺锤丝的收缩、使细胞染色体加倍的作用,对水稻幼穗诱导的愈伤组织细胞进行加倍,并分化出再生植株;创制出水稻同源四倍体新的种质材料,在此基础上选育水稻同源四倍体雄性不育三系材料,并实现水稻同源四倍体的三系配套,开展水稻同源四倍体杂种优势利用和四倍体杂交水稻选育研究,建立水稻同源四倍体杂种优势利用的新技术体系。这不仅有助于倍性水平杂种优势的开拓和利用,同时也将为我国新世纪“超级稻”育种研究开辟一条新的技术途径。 水稻幼穗诱导愈伤组织并分化成苗是一项成熟、简单的组织培养技术。本研究以普通二倍体水稻亲本为材料,用秋水仙素进行水稻的多倍体化诱导,创制同源四倍体水稻三系亲本材料并对其进行鉴定。多倍体化以秋水仙素诱导的愈伤组织培养为基础,研究不同秋水仙素浓度梯度和愈伤组织诱导培养基组合对诱导四倍体植株的影响。结果表明在MS+2,4 D 1.0mg/L+ KT0.2mg/L+ IAA0.2mg/L 和500mg/L的秋水仙素处理下,水稻愈伤组织染色体加倍(有最高的效率)效果较好,平均加倍频率可达25.26%,其中,材料CDR22和IR26诱导较易成功,加倍频率分别达到75%和26.5%;相对材料94109 1.3%加倍频率和冈46B 10.8%加倍频率,诱导率差异极显著。 对水稻四倍体材料进行了形态学鉴定结果表明,与二倍体水稻对照相比其株高、穗长、花粉育性等主要农艺性状,确定四倍体材料在穗长和千粒重两方面极显著提高,种子的长度和宽度也显著增长。对花粉育性鉴定,确认水稻四倍体不育系材料仍为不育,保持系材料自交和杂交可育,恢复系材料自交和杂交可育。对四倍体材料进行细胞形态、染色体数目等方面进行细胞学鉴定,经核型分析表明水稻四倍体材料具有48条染色体,是二倍体水稻的两倍。水稻四倍体材料根尖分生组织细胞与二倍体的根尖分生组织细胞相比,细胞体积、细胞核和核仁显著增大。四倍体三系材料在细胞有丝分裂中期均可规则排列在赤道板,并能均等地移向两极;后期观察中没有发现染色体分离滞后现象,分裂末期细胞能够形成大小相对均一的子细胞。水稻同源四倍体三系材料细胞分裂未见异常,植株生长发育正常。 从1996年至2006年,针对结实率、有效分蘖、着粒数和穗长等主要农艺性状,通过系谱选育的方法,对培育的同源四倍体水稻亲本材料进行了连续选择和改良,取得较好成效。表现为结实率的改良效果极佳,所有改良材料的平均结实率均呈上升趋势,如D237(29.70%→72.70%)、DTB(19.55%→53.21%)等。有效分蘖总体呈现上升趋势,但在不同的年份,如1998和2002存在较大的负向波动。部分材料改良效果明显,如D19B(5.87→13.50)、D什香 (7.00→12.00)等;同时一些材料如DTB和D明恢63虽然总体略有提高,但在不同的年份波动很大,因此存在较大改良阻力,原因还有待进一步研究。着粒数的改良上升趋势比较显著,除保持系的DTB之外,其余材料的平均着粒数有显著提高。穗长的改良阻力较大,虽然不同材料总体上有所提高,但效果并不显著,并且不同年份有较大负向波动(2001)。此外还对株高、剑叶长等性状也进行了选择,但效果不显著,原因有待进一步提高。同源四倍体材料产量相关性状遗传改良幅度不一致,保持系和恢复系间的遗传改良效果也存在差异。这为同源四倍体水稻的进一步利用打下了良好的基础。 籼稻和粳稻亚种间杂交及杂种优势利用的主要障碍就是其低的结实率。而同源四倍体杂交水稻的研究为提高杂交水稻的杂种优势利用创造了新的途径。本研究通过随机区组设计方案,挑选性状优良的二倍体水稻材料,包括雄性不育系,保持系和恢复系进行秋水仙素诱导加倍,从而获得同源四倍体水稻对应的三系材料。利用选育的优良水稻同源四倍体三系材料,配制7个杂交组合,杂交F1代与其恢复系亲本进行比较,用于计算超亲优势(HB);而杂交F1代与生产上大面积推广的二倍体杂交品种汕优63进行比较,用于计算杂种优势。结果显示,同源四倍体杂交水稻的超亲优势表现为:每株有效穗变化幅度为1.4%至105.9%,总粒数为0.5%至74.3%,每穗实粒数为17.6%至255.7%,结实率为9.6%至130.4%。这些农艺性状的改良使得这7个杂种F1的理论产量的超亲优势高达64.8%至672.7%。小区试验中四倍体杂交水稻组合T461A/T4002和T461A/T4193分别比二倍体对照汕优63提高46.3%和38.3%以上,除一个品种以外所有品种产量均接近或高于汕优63的产量。同源四倍体水稻强大的杂种优势表明,亚种间杂交育性低的问题可通过四倍体化及强化选择来解决。此外,同源四倍体杂交水稻器官的巨大性也是其产量提高的有利因素,水稻同源四倍体三系杂种优势利用研究具有一定的理论价值和商业生产潜力。 Rice is one of the major food crops, the improvement of the production and quality of it is an important thing related to the people's livelihood. On China's current national conditions, steadily increase of the rice yield based on the current level is still one of the most important goals in the next decade or even decades of agricultural production. According to the "super hybrid rice breeding" the strategic and rice breeding practice, improvement of the use of hybrid rice heterosis through continuous improvements in breeding technology is expected to get a higher level of rice yield, there are also a great yield potential can be exploited. However, there are also some difficulties to increase rice yield obviously and implement the goal of "super hybrid rice" based on the existing germplasm: Rice varieties in promotion are diploid, although there are many varieties, but their genome are from a comparatively single source; Meanwhile, the rice genome DNA are the least among the crops, it is difficult to exploit the development potential within the genome; Rice as C3 plants, photosynthetic efficiency is not high, it is one of the factors constraint rice yield. Therefore, seeking technological breakthroughs or innovative methods different from conventional means is the urgent needs to reach the target of "super hybrid rice". Using colchicine inhibit spindle contraction during cell division, double the cell chromosome, we induced callus cells from rice panicle to be doubled, and differentiated regeneration; we created a new autotetraploid rice germplasm material, and on that basis we bred male sterility three line autotetraploid rice materials, and the achieved the three line rice autotetraploid matchmaking, researched in autotetraploid rice heterosis usage and tetraploid hybrid rice breeding, constituted a new technology system of autotetraploid hybrid rice heterosis utilization. This not only helps the tetraploid rice heterosis exploration and use, but also inaugurates a new technical means for China in the new century "super rice" breeding research. We chose ordinary diploid rice as materials, using colchicine to induce the polyploidization, created the autotetraploid rice three-line materials and identified them. The polyploidization was based on the colchicine-induced callus tissue culture, and we experimented different colchicine concentrations and culture mediums to induce tetraploid plants, confirmed that the optimal concentration for inducement was 500 mg/L, the average induce rate was 25.26 %. Among all the materials, CDR22 and IR26 had higher induced rate; in contrary, 94109 and GANG46B had lower induced rate, the difference was significant. Autotetraploid materials was identified of both morphological and cytological, compared plant height, length of pollen sterility, and other major agronomic traits with a diploid rice as the control plant, identified that the autotetraploid materials had very significant advantages in ear length and thousand-grain weight, as well as the size of the seeds. Cytology identification included observation of the cell morphology, the number of chromosomes, and karyotype analysis on the autotetraploid materials confirmed that their chromosome number was 48, twice of the diploid rice. Mitoses in the three lines were common: chromosomes arrayed normally in metaphase and separated balanced into the two poles, chromosome moved without lagging in anaphase and daughter cells normally formed in telophase except one. It has been proved that tetraploid rice has normal meiosis as their diploid relatives, which usually including series of sub-phases as interphase, prophase I (five sub-phases), prophase II, metaphase I, II, anaphase I, II and telophase I, II. However, abnormal phenomena, such as formation of tetravalent, trivalent and univalent, chromosome lagging and so on, which would finally block meiosis. Configurations of chromosome in metaphaseⅠwere versatile in structure and form accept the bivalent. That condition varied in different strain, suggesting more complex paring configurations and more versatile genetic characters in tetraploid rice. All these abnormalities in meiosis contributed to low fertility of gamete and might consequently resulted in low seed setting. Successive selection and improvement on seed set, productive tiller per plant, total grains per panicle, panicle length and so on had been carried out from 1996 to 2006. The raise of seed sets was significant in both restorers and maintainers. Seed sets of some strains were improved more significantly than others, for example D237(29.70%→72.70%)、DTB(19.55%→53.21%)and et al.. Productive tiller per plant was improved to some extant. The tendency of improvement was rising on the whole but changed in some years such as 1998 and 2002. Part of the stains increased greatly, such as D19B(5.87→13.50)、Dshixiang (7.00→12.00) and so on, but some strains including DTB and Dminghui63 only increased little and decreased in some years by unknown reason. Total grains per panicle increased significantly and all strains except DTB increased. Improvement of panicle length termed to be hard. Different strains showed different capacities for improvement and floating existed in different years for example 2001. It has been proved that other agronomical traits including plant length, flag leaf length and so on could be improved but not significantly by selection also. In a word, agronomical traits could be raised by successive selection that is prerequisite for further utility of autotetraploid rice. Poor fertility is the main barrier for utilizing heterosis between the two rice (Oryza stiva L.) sub-species, indica and japonica. Recently, the development of autotetraploid hybrids (2n=4x=48) has been suggested as a new method for increasing heterosis in hybrid rice. Using standard experimental protocols, the elite diploid rice male sterile, maintainer, and restorer lines were colchine-doubled and autotetraploid counterparts were obtained. Seven resulting hybrids were analyzed for heterobeltiosis (HB), where the F1 was compared to the male parent, and the degree of heterosis, where the F1 was compared to the diploid commercial hybrid, Shanyou 63. The HB among the autotetraploid hybrids ranged from 1.4 to 105.9% for the productive panicles per plant, 0.5 to 74.3% for total kernels per panicle, 17.6 to 255.7% for filled kernels per panicle, and 9.6 to 130.4% for seed set. Improvements in these yield components resulted in the HB for kernel yield ranging from 64.8 to 672.7% among the seven hybrids. Hybrids T461A/T4002 and T461A/T4193 yielded 46.3 and 38.3% more, respectively than Shanyou 63, and all other hybrids but one yielded the same or more than Shanyou 63. The high heterosis for yield suggests that hybrid sterility between two rice sub-species may be overcome by using tetraploid lines followed by intensive selection. Also, the gigantic features of the autotetraploid hybrids may establish a plant structure able to support the higher yield.
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梗稻广亲和种质02428h中倒一节间伸长的性状由一对隐性基因(h基因)所控制,本实验证明了将h基因导入保持系与雄性不育系中均能得到遗传表达,促进倒一节的伸长。h基因与可育胞质及野败不育胞质作用后分别使穗伸出度达15cm,1.2cm,而相应珍汕97B及珍汕97A穗伸出度仅分别为1.7cm,-9.8cm。即h基因使穗伸出度分别提高8.8倍及1.2倍。因此,利用h基因有可能充分消除雄性不育系的包颈现象。The character of greatly elongated uppermost internode of wide compatible japonica rice 02428h iscontrolled by a recessive gene(b gene). The study shows that h gene could get genetic expression in maintaince andmale-sterile line(MS-line) and make the uppermost internode elonged. H gene interacts with the fertile and wild abortive male sterile cytoplasm make panicie neck exsert to 15cm and 1.2cm, while panicle neck exsertions of parents zhenshan 97B and A were 1.7cm and -9.8cm. So, h gene makes panicle necks exsert 8.8 and 1.2 times repectively. Its is possible to introduce h gene into male-sterile line so as to eliminate panicle endosure.
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用羟胺体外诱变质粒RP4,我们选择到一株温度敏感的菌落形成抑制突变体(temperature-sensitive colony-formation inhibition mutant,简称CFIts突变体)。其表型为在非允许温度时,在固体平板上不能由单个菌体繁殖为集落,类似于在非允许温度时,在固体平板上不能由单个菌体繁殖为集落,类似于文献报道的RP4的KiV基因表型。这种突变体初步认为是kor(KiV控制基因)的温度敏感性突变体,即由于kor蛋白产物的热变性导致KiV产物的过量或异常表达。生化分析研究表明:在非允许条件下,突变体的生长逐步受到抑制(其表现时倍增时间延长),大约繁殖两代后即全面抑制(菌数不再增加);此时宿主菌的DNA、RNA、蛋白质等大分子合成几乎全部下降,噬菌体产率也明显降低。此时状态是完全可逆的,即在全面抑制后的20小时内,仍可因放回允许温度培养而形成集落。据此抑制现象(KiV表型)的逐步发生性,大分子合成的全面抑制性和完全可逆性判断,很象是宿主菌株在紧急状态下的整体调节机制——严紧型反应。
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Time to flowering and maturity is an important adaptive feature in annual crops, including cowpeas (Vigna unguiculata (L.) Walp.). In West and Central Africa, photoperiod is the most important environmental variable affecting time to flowering in cowpea. The inheritance of time from sowing to flowering (f) in cowpeas was studied by crossing a photoperiod-sensitive genotype Kanannnado to a photoperiod-insensitive variety IT97D-941-1. Sufficient seed of F-1, F-2, F-3 and backcross populations were generated. The parental, F-1, F-2, F-3 and the backcross populations were screened for f under long natural days (mean daylength 13.4 h per day) in the field and the parents, F-1, F-2 and backcross populations under short day (10 h per day) conditions. The result of the screening showed that photoperiod in the field was long enough to delay flowering of photoperiod-sensitive genotypes. Photoperiod-sensitivity was found to be partially dominant to insensitivity. Frequency distribution of the trait in the various populations indicated quantitative inheritance. Additive (d) and additive x dominance (j) interactions were the most important gene actions conditioning time to flowering. A narrow sense heritability of 86% was estimated for this trait. This will result in 26 days gain in time to flowering with 5% selection intensity from the F-2 to F-3 generation. At least seven major gene pairs, with an average delay of 6 days each, were estimated to control time to flowering in this cross.
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Eukaryotic translation initiation factor 5A (eIF5A) is a protein that is highly conserved and essential for cell viability. This factor is the only protein known to contain the unique and essential amino acid residue hypusine. This work focused on the structural and functional characterization of Saccharomyces cerevisiae eIF5A. The tertiary structure of yeast eIF5A was modeled based on the structure of its Leishmania mexicana homologue and this model was used to predict the structural localization of new site-directed and randomly generated mutations. Most of the 40 new mutants exhibited phenotypes that resulted from eIF-5A protein-folding defects. Our data provided evidence that the C-terminal alpha-helix present in yeast eIF5A is an essential structural element, whereas the eIF5A N-terminal 10 amino acid extension not present in archaeal eIF5A homologs, is not. Moreover, the mutants containing substitutions at or in the vicinity of the hypusine modification site displayed nonviable or temperature-sensitive phenotypes and were defective in hypusine modification. Interestingly, two of the temperature-sensitive strains produced stable mutant eIF5A proteins - eIF5A(K56A) and eIF5A(Q22H,L93F)- and showed defects in protein synthesis at the restrictive temperature. Our data revealed important structural features of eIF5A that are required for its vital role in cell viability and underscored an essential function of eIF5A in the translation step of gene expression.
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Eukaryotic translation initiation factor 5A (eIF5A) is a protein that is highly conserved and essential for cell viability. This factor is the only protein known to contain the unique and essential amino acid residue hypusine. This work focused on the structural and functional characterization of Saccharomyces cerevisiae eIF5A. The tertiary structure of yeast eIF5A was modeled based on the structure of its Leishmania mexicana homologue and this model was used to predict the structural localization of new site-directed and randomly generated mutations. Most of the 40 new mutants exhibited phenotypes that resulted from eIF-5A protein-folding defects. Our data provided evidence that the C-terminal alpha-helix present in yeast eIF5A is an essential structural element, whereas the eIF5A N-terminal 10 amino acid extension not present in archaeal eIF5A homologs, is not. Moreover, the mutants containing substitutions at or in the vicinity of the hypusine modification site displayed nonviable or temperature-sensitive phenotypes and were defective in hypusine modification. Interestingly, two of the temperature-sensitive strains produced stable mutant eIF5A proteins - eIF5A(K56A) and eIF5A(Q22H,L93F)- and showed defects in protein synthesis at the restrictive temperature. Our data revealed important structural features of eIF5A that are required for its vital role in cell viability and underscored an essential function of eIF5A in the translation step of gene expression.
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Sweet sorghum figure as an alternative feedstock for ethanol production. The establishment of this culture in Brazilian production chain depends on the development of more productive and adapted cultivars. The aim of this study was to evaluate the general combining ability (GCA) of sweet sorghum lines and specific combining ability (SCA) of hybrid combinations as the agronomic and technological traits, and additionally to identify promising hybrid combinations for evaluation in advanced trials. Five restorer lines (R) and four male-sterile lines (A) were used in a partial cross diallel yielding 20 hybrids. The parental lines, hybrids and one check were evaluated in experiments carried out in a rectangular lattice design 5x6 with three replicates in two locations. The following traits were measured: flowering time, plant height, green mass yield, dry matter percentage, dry matter yield, juice extraction, total soluble solids content, sucrose content, purity, reducing sugars content, fiber content, sugars reducing total content, total recoverable sugars, hydrous ethanol, tons of per hectare, and ethanol production. There were differences between locations and genotypes for the traits. There was a significant effect of the genotype by environment interaction for most characters, except juice extraction, purity and reducing sugars content. There were a significant effect of GCA and SCA for most traits, indicating that additive and non-additive effects affect the phenotypic expression. Considering the effects of the GCA, the A line 201402B022-A, and R lines BRS 511, CMSXS643, and CMSXS646 were considered promising for exploration as parents in breeding programs of sweet sorghum in order to increase the ethanol production and the quality of the feedstock.The hybrids 201402B010-A x BRS 511, 201402B010-A x BRS 508, 201402B010-A x CMSXS646, 201402B022-A x BRS 511, 201402B022-A x CMSXS643, 201402B022-A x CMSXS646, 201402B022-A x CMSXS647 were the most promising for ethanol yield.
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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
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Pós-graduação em Agronomia (Horticultura) - FCA
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Pós-graduação em Agronomia (Proteção de Plantas) - FCA
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Pós-graduação em Biometria - IBB
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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)