Searching for New Sources of Pink Stem Borer Resistance in Maize (<Emphasis Type="Italic">Zea mays</Emphasis> L.)

The pink stem borer (Sesamia nonagrioides Lef.) is the main corn (Zea mays L.) pest in the Mediterranean area. Although, screening for resistance to this pest has been successful, the level of resistance shown by the most resistant varieties is not high. The objectives of the present work were: (i) the evaluation for pink stem borer resistance of the nontested inbred lines, field and popcorn materials, from the collection maintained at the Misión Biológica de Galicia and (ii) the study of the performance, under pink stem borer infestation, of inbreds selected for resistance to pink stem borer in hybrid combination. Forty four inbred lines nontested yet for resistant to pink stem borer along with five inbred lines previously reported as resistant to stem and ear attack, were evaluated in 2002 and 2003. In adjacent experiments, hybrids EP79× W552, EP77× B93, and A661× EP42 were tested along with a resistant hybrid, MEB531-Bt, and a susceptible hybrid, INRA 260. New sources of resistance to pink stem borer have been detected among inbred lines improving the level of resistance presented by previously tested inbreds. Some of these lines were successfully developed by pedigree selection for resistance to pink stem borer.     

Producing native seed of Stipa lagascae R. and Sch. inPresaharian Tunisia: high yields, high genetic variation orboth?

Genetic shifts are predicted to be greatest when highly diversematerial originating from an arid environment is bulk multiplied inirrigated and fertilised conditions. This is a key problem if largequantities of quality native seed are needed for restorationpurposes. In Presaharian Tunisia, we investigated two intermingledyield aspects of irrigated seed multiplication of Stipalagascae R. and Sch.: (1) nitrogen fertilisationand (2) selection for vegetative vigour. We set up twocontiguous nitrogen fertilisation trials, one withnon-selected seeds (from a nearby locality), and onewith selected seeds (48 clonal lines seeded in replicatedrows), and compared straw yields. Both selected and unselectedmaterial did not respond to more than 50 kg of nitrogenper ha. Unfertilised yields of both trials were similar,but the straw yield of fertilised selected material was more than100% higher than the straw yield of fertilised unselectedmaterial. In the trial with selected material, we collected straw andleaf yields per row, and scored each row for vigour and for phenologyduring three successive growing seasons. Genotypic differences inseed output and in mortality were large and mainly driven byvariation in potential straw for leaf ratios.The compromise between high yields and high genetic variation couldconsist of the following principles. (1) Create driergrowing conditions so that the actual straw forleaf ratio stays below the genotypic potential. (2) Useinstead fertilisation and selection to enhance seed yields.(3) Avoid bulk multiplication.     

Bambara groundnut (Vigna subterranea) seed quality in response to water stress on maternal plants

Farmers who still cultivate bambara groundnut (Vigna subterranea) rely on landraces and seed retained from previous harvests. Given that the crop is typically cultivated in semi-arid regions, seed quality of farmers' retained seed might be compromised due to water stress experienced by maternal plants during production. The aim of this study was to determine the effect of water stress on maternal plants on subsequent seed quality of bambara groundnut. A single bambara groundnut landrace was characterised into four distinct selections based on seed coat and speckling colour. Initial seed quality (viability and vigour) was evaluated prior to planting seed in a field trial under irrigated and rainfed conditions. Final yield and yield components were determined at harvest. Thereafter, seed quality (viability and vigour) of progeny of different landrace selections was evaluated. Yield was lower under rainfed than irrigated conditions. Overall, subsequent seed quality showed improvement from initial seed quality of the original seedlot. Seed viability was higher in seeds produced under irrigated than rainfed conditions. Seed quality of bambara groundnut may be reduced underwater-limited production conditions. Seed enhancement practices such as priming may assist farmers to achieve better emergence. In the long term, seed production should be done under optimum conditions in order to achieve high-seed quality.     

玉米种子超干燥方法及超干适应性研究

超干贮藏能在常温下长期保持种子发芽力,方便易行、高效低耗,但大粒种子超干燥困难.该试验以玉米杂交种农大108和自交系黄C为材料,用真空冷冻干燥机和烘箱干燥的方法进行超干处理,并对超干后的种子进行发芽测定,旨在探讨玉米一类大粒种子的超干适应性,筛选适宜的超干燥方法.结果表明,冷冻干燥机干燥较烘箱干燥速度快,能在较短时间内达到超干水平,且超干种子具有强的抗高温老化能力.但超干种子直接进行发芽,发芽率较低, 缓湿后发芽率明显提高.说明玉米种子可以进行超干贮藏,但必须进行缓湿处理后再播种.     

Germination of 151-year old <Emphasis Type="Italic">Acacia</Emphasis> spp. seeds

A collection of seeds from five Acacia species was made in Egypt in 1856. Since then, the seeds have been stored at room temperature in different Swedish museums. Due to the extreme longevity within the seeds of Acacia and related species, germination tests were performed on the now 151-year old seed. Seeds of two of the five species tested germinated. The first, Acacia farnesiana (L.) Willd., had two seeds germinate, and Acacia melanoxylon R. Br. ex Ait. f. had one seed germinate. In addition, DNA was extracted from the aged seed and DNA preservation was analyzed. Four of the tested species displayed well preserved DNA, whereas DNA from Albizia lebbeck (L.) Benth. showed signs of degradation. The 151-year longevity of the Acacia seeds is among the longest of dry-stored seeds reported. Several independent studies now report on extreme survival capacity for Acacia and related genera suggesting that these genera are suitable for studies on the characteristics of seeds with long storage performance. The results also demonstrate that herbaria and seed collections stored in museums and institutional depositories can be alternate sources of plants genetic material and should be given conservation attention.     

Viability of Oryza sativa L. seeds stored under genebank conditions for up to 30 years

Germination ability, equilibrium relative humidity (eRH), and moisture content of ‘control’ seed samples representing 183 rice accessions stored in the active (2–4 °C) and base (?10 °C until 1993, then ?20 °C) collections of the T. T. Chang Genetic Resources Center were determined after storage for 20.5–30.5 years. Germination of seeds that had been stored in the base collection was generally high (>70 %), whereas germination was more variable for seeds stored in the active collection. Samples with lower viability after storage in the active collection were likely to have lower viability after storage in the base collection. There were significant differences in the moisture content-eRH relationship of the seeds depending on whether the seeds had been stored in the active or base collection. Based on re-test data for regular seed samples regenerated in 1979–1980 and stored in the active collection for up to 31 years, estimates of the time for ability to germinate to fall to 50 % (p ₅₀) ranged from 54 to 997 years. For the same seed samples stored in the base collection for approximately 31 years, ability to germinate has been maintained and germination increased due to improved procedures. The ability to germinate of base collection samples was also generally higher than that of ‘safety duplicate’ samples of the same seed lots that had been sent to the National Center for Genetic Resources Preservation, USA in 1981 and stored at ?18 °C. This may have been due to uptake of moisture either during processing for dispatch or as a consequence of poor packaging material. The results are discussed in relation to long-term seed storage and genebank management.     

Association of a specific cationic peroxidase isozyme with maize stress and disease resistance responses, genetic identification, and identification of a cDNA coding for the isozyme

The presence of a pI 9.0 cationic peroxidase isozyme from milk stage pericarp of six susceptible and five resistant inbreds was correlated significantly with previously reported field data on percentage infection by Aspergillus flavus in the inbreds and their hybrids. The isozyme was constitutively expressed in some additional maize tissues and lines examined, and frequently induced by mechanical damage, heat shock, Fusarium proliferatum, and/or Bacillus subtilis in other lines tested. Native/IEF two-dimensional electrophoresis identified the isozyme as the previously genetically identified px5. A cDNA clone expressed in black Mexican sweet (BMS) maize cell cultures produced the pI 9.0 isozyme. In addition to potential use in marker-assisted breeding, enhanced expression of this cationic peroxidase through breeding or genetic engineering may lead to enhanced disease or insect resistance.     

Mode of inheritance of zinc accumulation in maize

The present study was undertaken to investigate the genetic nature of zinc (Zn) accumulation in the ear‐leaves of maize (Zea mays L.) plants. Fourteen inbred lines were evaluated. Out of which, five inbreds were chosen, two high Zn accumulators (Rg‐5 and Rd‐2), two low Zn accumulators (Rg‐8 and G‐307) and a moderate one (K‐64). These five parental inbreds were utilized to obtain all possible ten F1 hybrids (in one direction), 10 F2, 10 Bl, and 10 B2. All genotypes were planted and Zn was estimated at 50% tasseling stage. Zinc accumulation in the present genotypes was found to be genetically controlled and affected by additive genes. Significant values were obtained for the General Combining Abilities (GCA) but not for the Specific Combining Abilities (SCA) and the high Zn accumulating parents were the best general combiners. Four genes were found to be the minimum segregating factors in the (high x low) crosses for Zn accumulation.     

Maize (Zea mays L.) genetic factors for preventing fumonisin contamination

Fusarium moniliforme and Fusarium proliferatum are the most frequently isolated fungi from maize (Zea mays L.) in Spain. Both Fusarium species produce toxins potentially dangerous for animals and humans, the fumonisins being the most significant of those toxins. White maize is preferred for human consumption, and extra care should be taken to avoid kernel mycotoxin contamination. The objectives of this study were to identify and quantify kernel infection by Fusarium spp. and contamination by fumonisin on white maize hybrids, to search for white maize sources of resistance to infection by Fusarium spp. and mycotoxin contamination, and to preliminarily study the genetics involved in such resistances. Ten F(1) single crosses derived from a diallel mating design among five white maize inbreds were evaluated in a randomized complete block design with three replications in 2002 at two locations. Fusarium verticilloides and F. proliferatum were detected on kernels of white maize hybrids cultivated in northwestern Spain. No differences in fungal infection were found among maize genotypes, but differences in fumonisin contamination were significant and could be related, in part, to differences in husk tightness. Among the genotypes studied, general combining ability (GCA) effects were the most important for resistance to fumonisin contamination. Inbreds EP10 and EC22 showed the most favorable GCA effects for husk tightness and fumonisin content, and the cross between them, EP10 x EC22, had the most favorable specific combining ability (SCA) effect for husk tightness. Inbreds EP10 and EC22 showed favorable GCA effects for fumonisin contamination and husk tightness, and the cross EP10 x EC22 was the only one with an average fumonisin level below 1 mug/g. Although this should be confirmed with more extensive studies, white maize inbreds developed from white maize landraces could be sources of resistance to fumonisin contamination.     

Protein Distribution Pattern in Floury and Vitreous Endosperm of Maize Grain

Alpha‐amino nitrogen compounds of floury and vitreous parts of hand‐dissected endosperm from eight maize (Zea mays L.) inbred lines, representing a broad range of vitreousness (42–95%), were isolated as nonprotein nitrogen, albumin‐globulins, zeins, and true glutelins. The three protein classes averaged, respectively, 13, 48, and 35% of total nitrogen in floury endosperm, and 4, 79, and 15% of that in vitreous endosperm. For six inbreds, floury endosperm was richer in 27 kDa γ‐zein than vitreous endosperm; the reverse was found for an Argentine flint inbred (ARGL 256), and only traces of 27 kDa γ‐zein occurred in both floury and vitreous endosperm of inbred F113. Results were compared with protein distribution patterns reported in the literature of whole endosperm of wild‐type and mutant genotypes of maize, and with wild relatives of maize, Tripsacum, and teosintes. When percentage of salt‐soluble nitrogen increased from 2% (Tripsacum) to 22% (in double mutant Oh43o2;bt2), zeins decreased from 87 to 22%, and true glutelins increased from 11 to 57%. The pattern of whole endosperm of Zea perennis was very similar to that of the vitreous endosperm of line ARGL 256. The mean pattern for whole endosperm of six o2 inbred lines was identical to that of floury endosperm of eight wild‐type lines, consistent with a lack of synthesis of α‐zeins due to the mutation in the O2 gene.     

Preservation of seed viability during 25 years of storage under standard genebank conditions

Maintaining sufficient viability is critical to the sustainability of ex situ conserved seed collections. For this reason, accessions are regenerated when viability falls below a predefined threshold. Viability is monitored by determining the germination ability of accessions at predefined time intervals. Optimizing the frequency of these germination tests, in order to avoid waste of resources, is hampered by the scarce availability of data about seed longevity, particularly for material maintained under genebank conditions. Here we report on the analysis of nearly 40,000 germination test results collected for a wide range of crop species over a 25-years period by the centre for genetic resources, the Netherlands (CGN), where seeds of genebank accessions are dried to 3–7 % moisture content and stored for the long term under near-vacuum in aluminium foil bags at ?20 °C. The results indicate that seed viability is well maintained for the large majority of seed lots during the first 25 years after regeneration as only 3.3 % of the monitoring tests revealed below-threshold germination values. It is argued that the majority of these sub-standard seed lots are due to other causes than seed ageing, including dormancy problems and estimation error in germination testing. For material, maintained under the seed management procedures and storage conditions practiced by CGN, it is therefore recommended to delay the first germination monitoring tests to 25 years after regeneration.     

不同耐低磷玉米自交系生长发育特征研究

采用盆栽和田间试验方法研究了低磷胁迫对不同耐低磷玉米基因型全生育期生长发育的影响。结果表明,不同自交系在生长发育过程中对低磷胁迫的反应存在明显差异,在苗期耐低磷自交系植株干重和缺磷症状的相对值显著高于敏感自交系;到拔节始期,耐低磷自交系仍无缺磷症状,而敏感自交系症状加剧;进入生殖生长期后,耐低磷自交系孕穗期上部叶干重、茎干重受低磷影响的程度显著低于敏感自交系;至吐丝期,不同自交系间的耐低磷差异除了表现在植株干重上外,穗位高、雄花分枝数、功能叶面积也出现显著差异,低磷胁迫下耐低磷自交系这几个性状的相对值均显著高于敏感自交系;在成熟期,低磷胁迫下耐低磷自交系的相对产量较大,敏感自交系的相对产量较小。受低磷影响,敏感自交系的抽雄期、抽穗期、吐丝期和散粉期都显著延迟,而耐低磷自交系受影响较小或几乎不受影响。     

缺磷对不同耐低磷玉米基因型酸性磷酸酶活性的影响

【目的】酸性磷酸酶活性与土壤及植株体内有机磷的分解和再利用有着密切的关系。本研究以不同耐低磷玉米自交系为材料,研究低磷胁迫下玉米叶片、根组织内以及根系分泌酸性磷酸酶活性的变化及基因型差异,探讨酸性磷酸酶与玉米耐低磷之间的关系,以期更深入地了解玉米耐低磷的生理机制。【方法】以5个典型耐低磷自交系99180T、99239T、99186T、99327T、99184T和2个磷敏感自交系99152S、99270S为试验材料,采用营养液培养方法,设正常磷和低磷两种处理,分别于缺磷处理3、8和12 d时调查取样,测定地上部干重、根干重、叶片中无机磷(Pi)含量、根和地上部磷累积量、根系分泌APase活性以及叶片中APase活性,并于缺磷处理12 d测定根系内APase活性。【结果】1)缺磷使玉米地上部干重下降,根干重、根冠比增加,随着缺磷处理(3 d→8 d→12 d)时间的延长,根干重、根冠比增加幅度增大,且耐低磷自交系根干重增加幅度普遍大于敏感自交系。2)低磷条件下,玉米自交系磷吸收、利用效率存在基因型差异,耐低磷自交系99239T、99180T和99327T磷吸收效率较高,99186T和99184T磷利用效率高,敏感自交系99152S、99270S磷吸收和利用效率均较低。3)低磷处理使玉米自交系叶片无机磷(Pi)含量显著下降,耐低磷自交系99184T、99327T和99239T下降幅度较小,相对叶片无机磷含量较高。4)缺磷诱导玉米根系分泌的APase活性升高。耐低磷自交系99184T和99186T根系分泌APase活性升高幅度较大,其余3个耐低磷自交系未表现出明显优势。缺磷处理3 d、8 d,玉米根系分泌APase活性与磷累积量显著正相关,而12 d时相关性不显著;根系分泌APase活性与磷利用效率在缺磷处理12d时达显著正相关。说明玉米根系分泌APase活性与磷吸收、利用效率相关关系不稳定。5)缺磷处理12 d,各玉米自交系根组织内APase活性与根系分泌APase活性变化情况较一致,两者相关系数r=0.755(P0.05)。6)缺磷条件下各玉米自交系叶片组织内APase活性均有升高趋势,并表现出明显的基因型差异。缺磷处理8 d,耐低磷自交系99184T和99239T叶片组织内APase活性升高幅度最大,其次是99327T和99186T,99180T、99270S和99152S升高幅度较小;缺磷处理12 d,各玉米自交系叶片APase活性仍继续增加,99239T、99184T、99327T和99186T的相对APase活性均较高,99270S和99152S的相对APase活性较低。相关性分析表明,缺磷条件下玉米自交系叶片中相对APase活性与叶片中相对无机磷(Pi)含量显著正相关,与磷吸收、利用效率不显著相关。【结论】低磷诱导玉米叶片、根组织和根系分泌APase活性升高,根组织和根系分泌APase活性的大小与玉米耐低磷能力不完全相关,叶片APase活性与玉米耐低磷能力有较好的一致性。     

Reliable multiplication of seed for NERICA varieties of rice, <Emphasis Type="Italic">Oryza sativa</Emphasis> L.

A comparison of breeder seed from different seed sources produced guidelines for multiplication of NERICA (New Rice for Africa) varieties. Lower percentage detection of offtypes was noted with seed from a breeder than with seed from the Genetic Resources Unit (GRU) of the Africa Rice Center. The GRU grows breeder seed in the field as bulk populations of varieties and this seed is multiplied for the foundation seed. Breeders, however, always cultivate the variety in each family with several lines. In the comparison, each NERICA variety was cultivated as a single plant per hill for easy offtype detection in the field. Any offtypes or doubtful plants were recorded and immediately discarded. The breeder seed was not always genetically uniform because it was not yet fixed genetically and included mechanical seed mixture as well as natural mutants. Offtypes were classified into categories such as chlorophyll mutants, aberrations of plant height, plant types and panicle types, semi- or high sterility, awnness, apiculous color, leaf sheath color and excessively early plants. The causes of these offtypes were mainly (1) mechanical seed mixture, (2) segregation in genetically-unfixed material, (3) out-crossing with another variety, or (4) natural mutation. In 2006, we multiplied foundation seed using used home-saved seed from the 2005 crop harvested after removal of offtypes. The detection percentage of offtypes decreased compared with the previous year, but the number of semi- or high sterility plants increased significantly. This review confirms that breeder seed must be maintained with line culture within a family.     

Medium-term seed storage of 50 genera of forage legumes and evidence-based genebank monitoring intervals

Genebanks maintaining seeds for long-term genetic resources conservation monitor seed lots to detect early loss in viability. Monitoring is costly and depletes valuable seed. Three decades of genebank seed germination test results of diverse forage species from 50 legume genera in the International Livestock Research Institute’s medium-term store (circa 8 °C with 5% moisture content) were analysed to determine whether advice on seed monitoring intervals could be derived. Cumulative normal distributions were fitted by probit analysis for each seed lot and compared within each genus. Six patterns of within-genus variation were identified: no detectable trend in germination test results during storage (4 genera); detectable trends, but variable (positive to negative) amongst lots (5); consistent slope of loss in viability amongst lots (17); consistent slope of increase in ability to germinate amongst lots (21); common loss in viability amongst lots (2); common increase in ability to germinate amongst lots (1). Seed lot monitoring intervals for the medium-term store were derived for each of 19 genera with consistent loss in viability across seed lots: three genera provided comparatively rapid deterioration, five met the general expectations for a medium-term store (2–10 years’ maintenance of high viability), whilst 11 provided much better survival. Moreover, 26 further genera provided no evidence as yet of seed deterioration; of these, 22 improved in ability to germinate during storage indicating confounding of hardseededness with viability in germination tests.     

Characterisation of thirteen cowpea landraces from Umbria (Italy). Strategy for their conservation and promotion

Thirteen landraces of cowpea (Vigna unguiculata (L.) Walp. subsp. unguiculata cv. gr. Unguiculata) have been morphologically and agronomically characterised as part of a plan for safeguarding and protecting plant biodiversity typical of Lake Trasimeno (Umbria, central Italy). Sixty plants per accession were scored for plant vigour, growth habit, flower colour, days to flowering, days to first mature pod, pod length, total number of pods produced, seed colour pattern, 100 seed weight, total seed yield and virus susceptibility. With the exception of vigour and growth habit, all landraces appeared to be well differentiated from one another for all the characters considered. Strategies and problems related to their in situ conservation are discussed.     

利用SSR对60个玉米自交系进行杂种优势群划分初探

用SSR对60个玉米自交系的DNA进行分子标记和杂种优势群划分研究。利用14对SSR引物在供试材料中检测出57个等位基因变异,每对引物检测等位基因2～7个,平均为4．07个,多态信息量变化范围为0．389～0．832,平均为0．692。自交系间遗传相似系数变幅为0．058～0．756,UPGMA聚类分析表明,供试自交系可分为五个类群。利用这14对具有较高多态性信息量的引物,可以对供试材料进行初步鉴定。     

普通玉米和特用玉米的SSR标记聚类

利用SSR标记研究了64份玉米自交系（包括普通玉米和特用玉米自交系）的亲缘关系。用30对扩增带型稳定的SSR引物,从供试材料中检测出116个等位基因变异,每对引物检测出等位基因2~10个,平均3.87个。SSR引物的PIC介于0.306~0.893之间,平均多态性信息量为0.717。按UPGMA方法对供试自交系进行聚类,以遗传距离0.586为基准,可将材料划分为11类。64份自交系的SSR标记聚类分析表明,普通玉米与特用玉米糯玉米、甜玉米、爆裂玉米等并不能严格地各自划为一类,只有亲缘关系较近的特用玉米自交系聚在一起,有些特用玉米与普通玉米聚在一起,部分特用玉米具有丰富的普通玉米种质资源遗传背景。特用玉米自交系间的遗传距离与普通玉米自交系间的遗传距离差异达到0.01的极显著水平。此外,不同类型特用玉米间、特用玉米与普通玉米间的遗传差异较大,特用玉米在进化和选育过程中积累并保存了丰富的遗传变异。     

Low Level of Linkage Disequilibrium at the COMT (Caffeic Acid O-methyl Transferase) Locus in European Maize (<Emphasis Type="Italic">Zea mays</Emphasis> L.)

During recent decades, the whole plant yield of European maize (Zea mays L.) has increased substantially. However, during the same period of time cell wall digestibility, and consequently fodder quality, has decreased. Alleles for digestible cell walls have thus been lost, either during breeding for stalk standability or by genetic drift during breeding for grain yield. The brown-midrib 3 mutant (bm3) in maize (Z. mays L.), controlled by the caffeic acid O-methyl transferase (COMT) locus, has a positive influence on maize fodder quality. In this study, 42 European maize lines were used to evaluate the nucleotide diversity and linkage disequilibrium (LD) pattern across 2.3 kb of this locus. In agreement with what has been found for other maize loci, we found high diversity values (π = 0.00834), rapid LD decay even in an elite line sample, and indication of selection as well as of recombination events for seven site combinations. The diversity values differed slightly between inbreds of the Flint and Dent pools and most haplotypes were specific for one of the two heterotic groups. The polymorphisms identified at this single locus enable the construction of single nucleotide polymorphisms (SNP) and indel markers for haplotype identification and a rough discrimination of inbreds into Flint and Dent heterotic groups. The results reported here document the degree of allelic diversity in breeding materials and allow for targeted search of novel alleles in genetic resources.