Peanut stripe virus - a new seed-borne potyvirus from China infecting groundnut (Arachis hypogaea)1

A new virus, peanut stripe (PStV), isolated from groundnut (Arachis hypogaea) in the USA, induced characteristic striping, discontinuous vein banding along the lateral veins, and oakleaf mosaic in groundnut. The virus was also isolated from germplasm lines introduced from the People's Republic of China. PStV was transmitted by inoculation of sap to nine species of the Chenopodiaceae, Leguminosae, and Solanaceae; Chenopodium amaranticolor was a good local lesion host. PStV was also transmitted by Aphis craccivora in a non-persistent manner and through seed of groundnut up to 37%. The virus remained infective in buffered plant extracts after diluting to 10^-3, storage for 3 days at 20°C, and heating for 10 min at 60°C but not 65°C. Purified virus preparations contained flexuous filamentous particles c. 752 nm long, which contained a major polypeptide of 33 500 daltons and one nucleic acid species of 3·1 × 10⁶ daltons. In ELISA, PStV was serologically related to blackeye cowpea mosaic, soybean mosaic, clover yellow vein, and pepper veinal mottle viruses but not to peanut mottle, potato Y, tobacco etch, and peanut green mosaic viruses. On the basis of these properties PStV is identified as a new potyvirus in groundnut.     

Parsnip mosaic virus, a new member of the potato virus Y group

Parsnip mosaic virus (PMV) occurs commonly in parsnip in Britain and is transmitted after acquisition access periods of 2–5 min by the aphids Cavariella aegopodii, C. theobaldi and Myzus persicae. It was transmitted by manual inoculation of sap, infecting parsnip, chervil, coriander and carrot plants systemically, and causing local lesions without subsequent systemic infection in eight Chenopodium spp., Spinacia oleracea, Gomphrena globosa, and Toreniafournieri. It lost infectivity in Chenopodium quinoa sap after dilution to 10^-3–10^-4, heating for 10 min at 55–58 °C, or storage at room temperature for 7–10 days. Preparations partially purified by n-butanol or chloroform clarification, followed by acid precipitation and/or chromatography on columns of 2% agarose beads, contained filamentous particles, many of which were aggregated or fragmented. Preparations made with chloroform and without acid precipitation contained unaggregated particles of 755 nm normal length, with a sedimentation coefficient of 149 S. PMV did not react with antisera to any of fourteen other viruses with filamentous particles. The present cryptogram for PMV is */*: */*:E/E:S/Ap.     

Inheritance of resistance to groundnut rosette virus in groundnut (Arachis hypogaea L.)

A method of field screening groundnut seedlings for resistance to groundnut rosette virus (GRV), by means of which over 97% incidence was induced in rows of susceptible test plants, was developed at Chitedze Research Station in Malawi. Two GRV-resistant Virginia cultivars (RG 1 and RMP 40) were crossed with three susceptible cultivars, one from each of the Spanish (JL 24), Valencia (ICGM 48) and Virginia (Mani Pintar) botanical groups. Twelve F₁ reciprocal crosses and their F₂ and backcross generations were produced and the material screened in nurseries in 1985/86 and 1986/87. Seedlings raised from plants which did not become infected in the field were inoculated in the glasshouse in order to eliminate susceptible escapees. The numbers of diseased and healthy individuals in each population were subjected to χ² tests. In the majority of the F₂ populations a good fit was obtained for a ratio of one resistant to 15 susceptible plants, a ratio to be expected if resistance to GRV were determined by a pair of independent complementary recessive genes. This was further supported by data from backcross generations.     

花生成熟花粉的超微结构

花生（Arachis hypogaea L．）成熟花粉为二细胞型,具3个萌发沟,少数有4个。外壁呈蜂窝状。花粉壁由覆盖层、基粒棒、外壁内层及内壁构成。线粒体嵴密集、相互平行,脂体被粗面内质网包围。粗面内质网与外核膜相连,亦与线粒体相连。高尔基体甚少。营养核无核仁及染色质,与生殖细胞相连形成雄性生殖单位（male germ unit）。生殖细胞锤形、有壁,见一末端延伸成长尾状（长8μm）。细胞质含核糖体、线粒体、微管,未见体。在有些生殖细胞核内观察到具双层膜、少量嵴及深色内含物的球形结构,其米来源及本质尚不知,有待进一步研究确定。     

The biosynthesis of sulfoquinovosyldiacylglycerol: studies with groundnut (Arachis hypogaea) leaves

The biosynthetic pathway of sulfoquinovosyldiacylglycerol (SQDG) was investigated using groundnut (Arachis hypogaea) leaf discs and 35S-labeled precursors. [35S]SO4(2-) was actively taken up by the leaf discs and rapidly incorporated into SQDG. After 2 h, 1.5% of the [35S]SO4(2-) added to the incubation medium was taken up, of which 28% was incorporated into SQDG. The methanol-water phases of the lipid extracts of the leaf discs were analyzed for the 35S-labeled intermediates. Up to 2 h of incubation, cysteic acid, 3-sulfopyruvate, 3-sulfolactate, 3-sulfolactaldehyde, and sulfoquinovose (SQ) which have been proposed as intermediates [Davies et al. (1966) Biochem. J. 98, 369-373] were not labeled. Only a negligible amount of radioactivity was observed in these compounds after incubation for 4 h and more. Addition of sodium molybdate inhibited the uptake of [35S]SO4(2-) as well as its incorporation into SQDG by the leaf discs, suggesting that 3'-phosphoadenosine-5'-phosphosulfate may be involved in the biosynthesis of SQDG. Addition of unlabeled cysteic acid to the incubation medium enhanced the uptake of [35S]SO4(2-) but did not affect its incorporation into SQDG. 35S-labeled cysteic acid was taken up by the leaf discs and metabolized to sulfoacetic acid but not incorporated into SQ or SQDG. These results show that cysteic acid is not an intermediate in SQDG biosynthesis. [35S]SQ was taken up by the leaf discs and incorporated into SQDG in a time-dependent manner. [35S]Sulfoquinovosylglycerol was also taken up by the leaf discs but not incorporated into SQDG. It is concluded that SQDG is not biosynthesized by the proposed sulfoglycolytic pathway in higher plants. Though [35S]SQ was converted to SQDG, the rates are much lower compared to [35S]SO4(2-) incorporation, which suggests that a more direct pathway involving sulfonation of a lipid precursor may exist in higher plants.     

Root turnover of groundnut (Arachis hypogaea L.) in soil tubes

Five groundnut cultivars were grown in transparent tubes of pasteurized loam compost in growth-chamber conditions. Weekly tracings were made of all the roots visible through the walls of the tubes. White roots were assessed as living, and brown or decayed roots as dead; this correlated with microscopical assessments of root viability based on cytoplasmic staining with neutral red followed by plasmolysis.For all five cultivars, root laterals began to die 3–4 weeks after plants were sown. Death of root laterals progressed down the soil profile with time, while new roots were produced successively deeper from the extending taproot. The half-life of individual roots was calculated as 3.7–4.4 weeks for all cultivars, based on assessments of the roots that died up to plant maturity (14–20 weeks, depending on cultivar). At maturity, 73–83% of the cumulative length of root systems had died. The onset and rate of root death were not related to onset of flowering or pod-filling; instead, the peak times of root death at different distances down the root system were related to earlier (3–5 week) peak times of root production in those regions. The net result of root turnover was that, despite continued new root production, the maximum length of living (white) roots of each cultivar was recorded at 2–4 weeks after sowing. Death of the earliest formed root laterals was also observed in the first five weeks after sowing of groundnut in an experimental field plot in Malawi. Progressive root turnover is considered to be a normal feature of groundnut, perhaps representing an energy-economy strategy.     

Pod development of groundnut (Arachis hypogaea L.) in solution culture

Normal pods (containing seed) of groundnut (Arachis hypogaea L.) (cv. TMV-2) were successfully raised in darkened, aerated, nutrient solution, but not in the light. The onset of podding was evident 7 to 8 d after gynophores were submerged in the darkened nutrient solution. An examination of pods and submerged portions of gynophore surfaces by scanning electron microscopy showed the presence of two distinctly different protuberances: unicellular root-hair-like structures that first developed from epidermal cells of the gynophores and developing pods; and branched septate hairs that developed later from cells below the epidermal layer. The septate hairs became visible only after the epidermal and associated unicellular structures had been shed by the expanding gynophore and pods. Omission of Mn and Mg from the podding environment increased pod and seed weight, whilst omission of Zn reduced pod and seed weight.     

Genetic diversity of cucumber green mottle mosaic virus (CGMMV) infecting cucurbits

Cucumber green mottle mosaic virus (CGMMV), a well-known Tobamovirus, infects cucurbits across the globe. To determine its current status, molecular characterization, genetic recombination, gene flow and selection pressure, 10 districts from Punjab province of Pakistan were surveyed and a total of 2561 cucurbits samples were collected during 2019–2020. These samples were subjected to virus-specific double antibody sandwich-enzyme linked immunosorbent assay (DAS-ELISA) for the detection of CGMMV. The results revealed that viral disease was prevalent in all surveyed districts of Punjab with an overall 25.69% disease incidence. ELISA positive samples were further confirmed through RT-PCR and sequencing of coat protein (CP) cistron. Sequence analysis showed that the present studied CGMMV isolates have 96–99.5% nucleotide and 94.40–99.50% amino acid identities with those already available in GenBank. Phylogenetic analysis also revealed that understudied isolates were closely related with South Korean (AB369274) and Japanese (V01551) isolates and clustered in a separate clad. Sequence polymorphisms were observed in 663 bp of sequence within 31 CGMMV isolates covering complete CP gene. Total number of sites were 662, of which 610 and 52 sites were monomorphic and polymorphic (segregating), respectively. Of these polymorphic, 24 were singleton variable and 28 were parsimony informative. Overall nucleotide diversity (π) in all the understudied 31 isolates was 0.00010 while a total of 1 InDel event was observed and InDel Diversity (k) was 0.065. Haplotype diversity analysis revealed that there was a total 29 haplotypes with haplotype diversity (Hd) of 0.993458 in all the 31 isolates which provide evidence of less diversity among Pakistani isolates. The statistical analysis revealed the values 2.568, 5.31304 and 4.86698 of Tajima's D, Fu, & Li’s F* and D*, respectively, which witnessed the population of CGMMV was under balanced selection pressure.     

Activities of Various Peptidases in Cotyledons of Germinating Peanut (Arachis hypogaea)

The activities of several carboxy- and aminopeptidases were assayed in extracts prepared from the cotyledons of resting and germinating peanut seeds as well as from growing and fully differentiated peanut leaves. Carboxypeptidases acting on two carbobenzoxydipeptides Z-Phe-Ala and Z-Ala-Phe at pH 5.2 showed minimal activities in “resting” cotyledons, and only slight increases occurred during 7-day germination at 28°C. In peanut leaves the corresponding activities were quite high, about 20- and 6-fold compared to “germinating” cotyledons. Peptidases acting on Leu-Tyr at pH 8.6 and on Ala-Gly at pH 7.8 were highly active in resting cotyledons, and the activities remained essentially constant during germination; corresponding activities in leaves were much smaller (about 15–25% of those in cotyledons). “Naphthylamidases” hydrolyzing the β-naphthylamides of Phe, Leu, and Arg at pH 7.2, were also highly active in resting cotyledons; during germination the first activity stayed at a constant level while the other two decreased progressively. Leaves showed relatively high activities on Phe-bT-NA and Leu-β-NA but only minimal activity on Arg-β-NA. It is tentatively concluded that the peptidases acting on Leu-Tyr and on Ala-Gly as well as the naphthylamidases function in the mobilization of the reserve proteins of peanut cotyledons during germination. The carboxypeptidases, in contrast, do not seem to play a major role in this process.     

22Na influx is significantly lower in salt tolerant groundnut (Arachis hypogaea) varieties

Distinct varieties differing in salt tolerance were initially identified from two separate green house experiments using two systems; solution as well as soil culture. The first screening involved a diverse group of 27 cultivars. Several physiological traits; Chlorophyll Stability Index (CSI), Salt Tolerance Index (STI) and ion content were determined to screen the cultivars for differences in salt tolerance using solution culture in the first experiment. A set of six varieties (three tolerant and three susceptible) were selected from this experiment and then subjected again to salt stress adopting a natural soil system in the second experiment which involved a screening approach essentially similar to that of the first experiment. In the third experiment using two distinct cultivars differing in salt tolerance selected from experiment II, ²²Na influx rate was determined in the root and shoot at the end of a 24 h salt imposition in Hoagland’s nutrient system containing 180 KBq of ²²Na. The results suggested that there were distinct differences in ²²Na influx rate into root and concurrently in the shoot. The salt tolerant Spanish improved and one of the moderately tolerant Trombay variety TAG 24, showed good regulation of ²²Na influx resulting in low ²²Na concentration. The salt susceptible variety JSP39 had nearly 7–8 fold higher root ²²Na content as compared to the tolerant and moderately tolerant cultivars. The results have highlighted the importance of Na exclusion as an important determinant of salt tolerance in groundnut.     

花生种子耐脱水力的形成与可溶性糖累积的关系

花生胚轴的耐脱水力在40－65DAP的发育后期逐渐增加,同时胚轴还原性糖／非还原性糖比值下降。缓慢干燥可同时诱导35DAP胚轴累积蔗糖与寡糖（水苏糖与棉籽糖）;外源ABA及高渗处理可诱导35DAP离体胚轴累积蔗糖,但不能累积寡糖（水苏糖）。耐脱水胚轴可溶性糖成分的模拟混合物可在水活度0．32及零上温度进入玻璃态;而不耐脱水胚轴的可溶性糖模拟混合物仅在零下温度进入玻璃态。模拟实验证明在干燥状态下可溶性糖与花生2S蛋白结合,并消除了2S蛋白的干燥结晶。     

Molecular tagging of a rust resistance gene in cultivated groundnut (Arachis hypogaea L.) introgressed from Arachis cardenasii

Rust is a serious and the most prevalent groundnut disease in tropical and subtropical growing regions of the world. A total of 164 recombinant inbred lines derived from resistant (VG 9514) and susceptible (TAG 24) cultivated groundnut parents were screened for rust resistance in five environments. Subsequent genotyping of these lines with 109 simple sequence repeat (SSR) markers generated a genetic linkage map with 24 linkage groups. The total length of the linkage map was 882.9 cM with an average of 9.0 cM between neighbouring markers. The markers pPGPseq4A05 and gi56931710 flanked the rust resistance gene at map distances of 4.7 cM and 4.3 cM, respectively, in linkage group 2. The significant association of these two markers with the rust reaction was also confirmed by discriminant analysis. The informative SSR markers classified rust-resistant and susceptible groups with 99.97% correctness. The SSR markers pPGPseq4A05 and gi56931710 were able to identify all the susceptible genotypes from a set of 20 cultivated genotypes differing in rust reaction. Tagging of the rust resistance locus with linked SSR markers will be useful in selecting the rust resistant genotypes from segregating populations and in introgressing the rust resistance genes from diploid wild species.     

Characterization of a New Strain of Capsicum chlorosis virus from Peanut (Arachis hypogaea L.) in China

A new strain of Capsicum chlorosis virus (CaCV) from peanut (Arachis hypogaea L.) in China, designated CaCV‐CP, was characterized. CaCV‐CP causes yellow spots and necrosis on the leaves of affected peanut plants. Of 31 plant species inoculated mechanically, 24 were susceptible to this strain. Quasi‐spherical virions were present in ultrathin section of diseased leaves. The complete sequence of S RNA of CaCV‐CP consisted of 3399 nucleotides (nts). The NSs and N genes of the virus contained 1320 nts and 828 nts, respectively; these two open reading frames were in an ambisense arrangement. The N gene of CaCV‐CP shared 84.7–86.4% and 92.4–93.1% identity with that of CaCV strains from Thailand and Australia at nt and amino acid levels, respectively (the GenBank accession number of the sequence reported in this study is DQ355974 ).     

Response of groundnut (Arachis hypogaea L) toRhizobium inoculation in the field: Problems and prospects

Summary A successfulRhizobium inoculant strain needs to be more competitive than native soil rhizobia in forming nodules and to be effective in fixing nitrogen. Persistence of the strain from one season to the next will be an added advantage, since it eliminates the need to inoculate every season. Earlier studies indicated that groundnut,Arachis hypogaea, seldom responds toRhizobium inoculation in soils already containing rhizobia which nodulate groundnut. However, the results of seven years of research at ICRISAT have opened up the prospect of inoculating groundnut in such soils. These experiments indicate that inoculating with sufficient numbers of an effectiveRhizobium strain, NC 92, applied as a liquid slurry below the seed, increased yields of certain groundnut cultivars. Similar results were reported from other research centres in India. Inoculation with NC 92 for two consecutive seasons increased the proportion of nodules formed by this strain, from 25–32% in the first season to 41–54% in the second season, indicating that this strain can persist in the field for the following season. This paper also discusses results of experiments relating to host cultivar specificity and some possible problems in applying this information to farmers' fields. Strain NC 92 also produces a siderophore, an iron chelating compound, which may help in the iron nutrition of the plant.

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Resumen Respuesta del maní (Arachis hypogaeaL.)a la inoculación con Rhizobiumen el campo. Problemas y perspectivas.La inoculación conRhizobium para tener exito debe de realizarse con una cepa que sea competitivamente mejor que las cepas nativas del suelo en cuestión, en relación a la habilidad para formar nódulos y a la eficacia en la fijación de nitrógeno. La persistencia de la cepa de una temporada para otra es una caracteristica deseable ya que elimina la necesidad de reinocular cada temporada. Estudios previos indicaban que el maní (Arachis hypogaeae) raras veces responde a la inoculación conRhizobium en suelos que ya contienenRhizobium especificos. Sin embargo los resultados de siete años de investigaciones en ICRISAT han abierto nuevas perspectivas para la inoculación del maní en dichos suelos. Esta experiencias indican que la inoculación, en cantidades suficientes de una cepa deRhizobium eficaz (NC 92) aplicada en forma de liquido viscoso bajo la semilla incrementa el rendimiento de ciertos cultivars de mani. Resultados semejantes se han observado en otros centros de investigación en India.La inoculación de la cepa NC 92 durante dos años consecutivos incrementó la proporción de nódulos formados desde 25–32% en la primera temporada hasta 41–54% en la segunda indicando pues la persistencia de dicha cepa en el suelo de una a otra temporada. Este trabajo también evalua los resultados de investigaciones relacionando la especificidad del inóculo con la posible problemática que conlleva la aplicación de esta informacion por el agricultor.

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Résumé Réponse de l'arachide (Arachis hypogeaL.)à l'inculation de Rhizobiumdans les champs. Problèmes et perspectives.Pour être couronnée de succès, une souche deRhizobium inoculée doit supplanter les rhizobiums natifs du sol en ce qui concerne la formation de nodules et doit fixer l'azote efficacement. La persistance de la souche d'une saison à l'autre, rendant inutile la répétition saisonnière de l'inoculation, est un avantage supplémentaire. Des études antérieures ont montré que l'arachide (Arachis hypogea) ne répond que rarement à l'inoculation dans des sols contenant déjà des rhizobiums capables de noduler cette plante. Cependant, les résultats de sept années de recherches éffectuées à l'ICRISAT ont ouvert des possibilités quant à l'inoculation dans ce type de sols. Ces expériences montrent qu'une bouillie liquide d'une souche efficace deRhizobium (NC 92), répandue en quantité suffisante sous les graines, augmente le rendement de certains cultivars d'arachide. Des résultats similaires ont été rapportés par d'autres centres de recherches en Inde. L'inoculation de NC 92 pendant deux saisons consécutives a augmenté la proportion des nodules formés par cette souche de 25 à 32% pour la première saison, et de 41 à 54% pour la seconde, ce qui indique que cette souche peut persister dans le sol d'une saison à l'autre. Dans cet article sont également discutés les résultats d'expériences concernant la spécificité de l'hôte et quelques problèmes concernant l'utilisation de ces informations par les agriculteurs. La souche NC 92 produit aussi un sidérophore, agent complexant du fer pouvant jouer un rôle dans le métabolisme de ce métal chez la plante.

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Submitted as ICRISAT J. A. No. 475.     

花生根瘤菌群体遗传多样性和系统发育研究

利用16S rRNA RFLP,16S rRNA序列分析和16S－23S IGS PCR RFLP技术对43株花生根瘤菌和来自其他种属的15个参比菌株进行了群体遗传多样性和系统分析。16S rRNA PCR RFLP分析结果表明,所有供试花生根瘤菌均属于慢生根瘤菌属,在系统发育上与B.japonicum的亲缘关系最近,具有相同的16S rRNA RFLP基因型,而与B.elkanii相对较远。16S rRNA 序列分析结果表明,供试花生根瘤菌在系统发育上更接近于B.liaoningense,序列间差异小于1％,而B.liaoningense在系统发育上与B.japonicum相距很近,其序列间差异小于1％,16S－23S rRNA IGS RFLP分析结果表明,尽管花生根瘤菌与B.japonicum和B.elkanii的亲缘关系很近,但在71％的相似性水平上供试花生根瘤菌仍各自聚为一群,并可进一步分为A、B、C和D4个亚群,该分群还明显反映了地理因素对群体遗传多样性和系统发育的影响。     

Variation and inheritance of the arachin polypeptides of groundnut (Arachis hypogaea L.)

Summary Variation in the arachin polypeptides of groundnut genotypes was observed by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). Three regions could be observed on the electropherogram. Region 1, corresponding to conarachin, did not show any variation; region 2, consisting of arachin acidic subunits, showed variation; region 3, containing the arachin basic subunits, did not show any variation. There are four varietal classes of arachin polypeptide patterns: class A comprised three acidic subunits of arachin of molecular weights 47.5, 45.1 and 42.6 kd and a basic subunit of 21.4 kd; class B, with three acidic subunits of molecular weights 47.5, 45.1 and 41.2 kd and a basic subunit of 21.4 kd; class C of an additive pattern of class A and class B; class D, of two acidic polypeptides of 47.5, 45.1 kd and the basic 21.4 kd subunit. Of the 90 genotypes studied, 73% belong to class A, 15% to class B and 6% each to class C and D. Analysis of F₂ seeds from a cross between class A and class B genotypes showed that the two polypeptides (42.6 kd and 41.2 kd) are coded by nonallelic genes and also revealed that class C and class D patterns arose as a result of hybridisation between class A and class B. A. monticola, the progenitor of A. hypogaea, showed a pattern similar to the additive pattern of class A and class B while some diploid Arachis species had the 41.2 kd polypeptide. Based on arachin polypeptide patterns the probable origin of A. hypogaea has been suggested.     

花生转基因研究进展

花生是世界上重要的油料和经济作物之一,是人们生活的植物脂肪和蛋白质来源。现代生物技术的不断发展为花生育种和种质创新提供了新的技术手段, 它可以直接将来自不同种属的异源目的基因插人到花生基因组, 使花生表达目标性状, 实现花生品种的遗传改良。近年来, 国内外花生转基因研究取得了重大进展。文章综述了花生转基因在抗虫、抗病、抗非生物逆境和品质改良等方面的最新进展,并总结了近年来人们对农杆菌介导法、基因枪法和不依赖组织培养的转化法等主要的花生遗传转化方法的改进和探索。     

花生根部性状的遗传分析

利用花生RIL群体,分析了11个花生根部性状的遗传力,估算基因对数及性状间的相互关系,根据偏度系数(g1)和峰度系数(g2)估算控制性状的基因互作情况。结果表明:11个性状都是受多基因控制的数量性状,在RIL群体中基因型间的差异均表现为连续变异和明显的超亲分离。侧根干重的遗传力最高达0.60,其次是侧根鲜重,为0.58,而其他性状的遗传力均较低。控制主根长性状的多基因间存在互作,互作方式为重叠作用;控制主根粗(3cm)性状的基因间也存在一定的重叠作用,但是作用不明显;控制其他性状的基因都存在互作,表现为互补作用,但互补作用的强弱有差异。主根粗(1cm)、主根粗(3cm)、主根干重、主根鲜重、侧根干重和侧根鲜重之间都显著或极显著相关;根体积与主根粗(1cm)、主根粗(3cm)、侧根干重和侧根鲜重显著或极显著相关。