Effects of desiccation on peanut (Arachis hypogaea L.) seed protein composition

Response of peanut to desiccation was studied by monitoring changes in the seed protein content and composition during a 14 day desiccation period using a combination of electrophoretic and immunochemical techniques. Following desiccation, the protein content of ‘white’ (most immature) and ‘orange’ seed increased, while that of the ‘brown’ (mature) seed was not affected. Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) showed no major qualitative differences in the protein composition during desiccation of the samples. However, immunoblotting with anti-dehydrin antisera revealed the presence of several new proteins in the desiccated samples compared with the controls. One of the dehydrin-like proteins, band ‘c’ was found to be related to water-stress, while the other proteins appeared to be the storage proteins accumulated as the seed matured in vitro. Capillary electrophoresis (CE) showed major changes in the protein quantity and quality of ‘white’ seed during the 0–14 days of desiccation. In contrast, in the ‘orange’ and ‘brown’ seeds changes in protein composition were less significant. Results indicated that there are several dehydrin-like proteins expressed in peanuts, however, not all of them are related to water stress.     

Shoot organogenesis from cultured seed explants of peanut (Arachis hypogaea L.) using thidiazuron

Summary Thidiazuron (TDZ) was utilized to induce adventitious shoot formation from the hypocotyl region of cultured seed explants of peanut (Arachis hypogaea L.). Excision of the radicle from seed explants was more stimulatory to shoot initiation than removal of the epicotyl alone. Removal of both the radicle and the epicotyl from seeds resulted in a 37-fold increase in the frequency of shoot production when compared to intact seeds. Half seed explants with epicotyl and radicle removed produced the greatest number of shoots per explant. Explants from mature seeds were more responsive to TDZ than immature seed-derived explants. A 1-wk exposure to 10 μM TDZ was sufficient to stimulate the initiation of adventitious shoots that subsequently developed into plants. High frequency of shoot initiation was readily induced in a variety of genotypes ofA. hypogaea and a wild peanut (A. glabrata). Plants regenerated from shoots induced by TDZ were phenotypically normal and fertile.     

Response of two peanut (Arachis hypogaea L.) cultivars to boron and calcium

The response of two peanut cultivars (Tainan 9 and SK 38) to applications of six boron (B) rates (H₃BO₃ at 0, 0.12, 0.25, 0.5, 1 and 2 kg B ha^–1) at two calcium levels [nil (-Ca) or CaSO₄ at 100 kg Ca ha^–1 (+Ca)] to a B-deficient Oxic Paleustult was studied in a pot experiment. Without added Ca, both cultivars had low seed yields and gave only small responses to B. Similarly, without added B, both cultivars had low seed yields and did not respond to Ca. But, with added Ca or B, they responded strongly to B and Ca, respectively. In both cultivars, deficiencies of Ca or B depressed seed dry weight by depressing seed size by over 75%. Boron deficiency further depressed seed dry weight by decreasing the number of seeds per plant by decreasing the number of seeds per pod in Tainan 9, and the number of pods per plant in SK 38. Seed dry weight was depressed more than pod dry weight, so that both Ca and B deficiencies severely depressed the shelling %. With added Ca, tainan 9 responded to lower levels of B than SK 38, reaching maximum seed dry weight at 0.12 kg B ha^–1. At this low level of B. SK 38 yielded only half the seed dry weight of Tainan 9. But SK 38 continued to respond to increasing levels of B to 2 kg ha^–1, producing a maximum seed yield 40% higher than Tainan 9. The results indicate that where all other nutrients are adequate, SK 38 will yield better than Tainan 9 on soils with high B, but worse on soils with low B. The implications of these findings for the selection of peanut cultivars are discussed.     

Quantification of the geocarposphere and rhizosphere effect of peanut (Arachis hypogaea L.)

Roots and pods of field-grown peanut (groundnut) (Arachis hypogaea L.) were sampled at the R3, R5, and R7 developmental stages and examined in comparison to root- and pod-free soil for microbial population densities to assess the geocarposphere and rhizosphere effects. G/ S (no. geocarposphere microorganisms/no. soil microorganisms) and R/S (no. rhizosphere microorganisms/no. soil microorganisms) ratios were calculated for total fungi,Asperigillus flavus, spore-forming bacilli, coryneform bacteria, fluorescent pseudomonads, and total bacteria isolated on low- and high-nutrient media. A clear geocarposphere effect was evidenced by increased population densities of bacteria and fungi associated with developing pods compared to soil. G/S and R/S ratios were generally greater than 1.0 for all groups of microorganisms except bacilli. G/S ratios were greater for total bacteria than for total fungi at two of the three sample times, suggesting that bacteria were stimulated more than fungi in the zone around developing pods. In contrast, R/S ratios, were higher for total fungi than for total bacteria at two of three sample times. The preferential association of fungi and bacteria with early developmental stages of the pod indicates that some microorganisms are particularly well adapted for colonization of the peanut geocarposphere. These microorganisms are logical candidates for evaluation as biological control candiates forA. flavus.     

Fungal populations in the rhizosphere of peanut (Arachis hypogaea L.)

Summary A comparison of fungal populations in the rhizospheres of eight varieties of peanut grown in a red lateritic soil amended with farmyard manure was made by the dilution-plate technique. There was a marked increase in fungi in the rhizospheres of TMV 2, TMV 4, Pollachi Red and EC 1698, the increase was smaller in Spanish Improved and RS 1 while very little rhizosphere effect was shown by TMV 3 and Pondicherry 8. Age of the plant had a significant influence on numbers of fungi in the rhizosphere. High R/S ratios were obtained when the plants were 30 days old, at which time attained maximum vegetative growth and started to flower. The ratios gradually decreased after that age until the plants were three months old when there was again a small increase. This later rise in fungal populations is interpreted to be due to an increase in microbial activity around dead or senescent roots. No correlation could be established between numbers of root nodules produced by a variety and its rhizosphere effect. Preferential stimulation of certain fungi in the rhizosphere of some of the varieties was noticed.     

去胚轴对花生子叶肽链内切酶和贮藏蛋白质降解的影响

从离体子叶与连体子叶在水中培养一段时间后的比较,看到它们之间在肽链内切酶活性和盐溶蛋白及花生球蛋白降解上的差异并不大,这表明去除胚轴对子叶肽链内切酶活性和贮藏蛋白降解的影响很轻微。亚胺环己酮(蛋白质合成抑制剂)不能完全抑制离体子叶肽链内切酶活性的提高,子叶的大部分大分子贮藏蛋白同样被降解。这表明,在花生种子萌发过程中降解大部分贮藏蛋白的子叶肽链内切酶并非全部是在种子萌发时新合成的,子叶贮藏蛋白降解和肽链内切酶活性基本不受胚轴调控,子叶与胚轴之间在调控关系上可能是一种新的调节类型。     

Phylloplane bacteria increase seedling emergence, growth and yield of field-grown groundnut (Arachis hypogaea L.)

AIM: To isolate and characterize groundnut-associated bacterial isolates for growth promotion of groundnut in field. METHODS AND RESULTS: Three hundred and ninety-three groundnut-associated bacteria, representing the geocarposphere, phylloplane and rhizosphere, and endophytes were applied as seed treatment in greenhouse. Maximum increase in plant biomass (up to 26%) was observed following treatment with a rhizosphere isolate identified as Bacillus firmis GRS 123, and two phylloplane isolates Bacillus megaterium GPS 55 and Pseudomonas aeruginosa GPS 21. There was no correlation between the production of L-tryptophan-derived auxins and growth promotion by the test isolates. Actively growing cells and peat formulations of GRS 123 and GPS 55, and actively growing cells of GPS 21, significantly increased the plant growth and pod yield (up to 19%) in field. Rifampicin-resistant mutants of GRS 123 and GPS 21 colonized the ecto- and endorhizospheres of groundnut, respectively, up to 100 days after sowing (DAS), whereas GPS 55 was recovered from both the habitats at 100 DAS. CONCLUSION: Seed bacterization with phylloplane isolates promoted groundnut growth indicating the possibility of isolating rhizosphere beneficial bacteria from different habitats. SIGNIFICANCE AND IMPACT OF THE STUDY: Identification of phylloplane bacteria as effective plant growth-promoting rhizobacteria (PGPR) broadens the spectrum of PGPR available for field application.     

Effect of microspore stage and media on anther culture of peanut (Arachis hypogaea L.)

This study was designed to study the effects of stage of microspore development and culture medium on androgenic response in peanut (Arachis hypogaea L.). Anthers of various developmental stages were cultured for 7 days, then fixed and observed cytologically. Three sets of media, involving different basal media, growth regulators, sucrose levels and glutamine concentrations, were tested. In all experiments, the stage of development of the microspores at the time of culture was highly significant. The early uninucleate microspores stage was identified as producing the highest anther response rating. The effect of media was nonsignificant in all experiments. However, the stepwise modification of the media through the course of the study resulted in an almost 8 x increase in anther response rating. Numerically, the best media tested was N₆ basal medium with 1 mg 1^-1 NAA, 0.1 mg 1^-1 BA, 5.5% sucrose, and 3.5 g 1^-1 glutamine. While no haploids were obtained, four-nucleate cells were observed, indicating the potential in peanuts for an androgenic reponse.     

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。     

Evaluation of peanut (Arachis hypogaea L.) leaflets from mature zygotic embryos as recipient tissue for biolostic gene transfer

Leaflets from mature peanut embryos are a useful recipient tissue for biolistic DNA transfer. Fertile plants were regenerated from leaflets from genotypes representing all botanical types of peanut. Regeneration frequency was strongly influenced by genotype. NPT II and GUS chimaeric gene fusions, driven by the CaMV 35S promoter, were expressed transiently following biolistic delivery to unexpanded leaflets. Bombardment conditions affecting transient expression frequency were determined using a prototype of the Bio Rad PDS 1000/He helium-powered particle acceleration apparatus. Stably transformed calli were derived routinely from leaflet tissue bombarded with the NPT II gene and subsequently cultured on kanamycin. Several plants have been regenerated from treated explants under kanamycin selection. Thus far, none of these has been stably transformed. The occurrence of escapes suggests that kanamycin is an inefficient selective agent for the recovery of transgenic peanuts from this explant. Experiments designed to regenerate plants using published regeneration protocols from stably transformed calli, devoid of primary explant tissue, have been unsuccessful.     

Diversity and compatibility of peanut (Arachis hypogaea L.) bradyrhizobia and their host plants

A high degree of genetic diversity among 125 peanut bradyrhizobial strains and among 32 peanut cultivars collected from different regions of China was revealed by using the amplified fragment length polymorphism (AFLP) technique. Eighteen different peanut bradyrhizobial genotypes and six peanut cultivars were selected for symbiotic cross-inoculation experiments. The genomic diversity was reflected in the symbiotic diversity. The peanut cultivars varied in their ability to nodulate with the strains used. Some cultivars had a more restricted host range than the others. Also the strains displayed a range of nodulation patterns. In yield formation there were clear differences between the plant cultivar/bradyrhizobium combinations. There was good compatibility between some peanut bradyrhizobial strains and selected cultivars, with inoculation resulting in well-nodulated, high-yielding symbiotic combinations, but no plant cultivar was compatible with all strains used. The strains displayed a varying degree of effectiveness, with some strains being fairly effective with all cultivars and others with selected ones. The AFLP genotypes of the strains did not explain the symbiotic behavior, whereas the yield formation of the plant cultivars was more related to the genotype. It is concluded that to obtain optimal nitrogen fixation efficiency of peanut in the field, compatible plant cultivar-bradyrhizobium combinations should be selected either by finding inoculant strains compatible with the plant cultivars used, or plant cultivars compatible with the indigenous bradyrhizobia.     

Diversity characterization and association analysis of agronomic traits in a Chinese peanut （Arachis hypogaea L.） mini-core collection

Association mapping is a powerful approach for exploring the molecular basis of phenotypic variations in plants. A peanut （Arachis hypogaea L.） mini-core collection in China comprising 298 accessions was genotyped using lo9 simple sequence repeat （SSR） markers, which identified 554 SSR alleles and phenotyped for 15 agronomic traits in three different environments, exhibiting abundant genetic and phenotypic diversity within the panel. A model-based structure analysis assigned all accessions to three groups. Most of the accessions had the relative kinship of less than o.05, indicating that there were no or weak relationships between accessions of the mini- core collection. For 15 agronomic traits in the peanut panel, generally the Q ＋ K model exhibited the best performance to eliminate the false associated positives compared to the Q model and the general linear model-simple model. In total, 89 SSR alleles were identified to be associated with 15 agronomic traits of three environments by the Q＋K model-based association analysis. Of these, eight alleles were repeatedly detected in two or three environments, and 15 alleles were commonly detected to be associated with multiple agronomic traits. Simple sequence repeat allelic effects confirmed significant differences between different genotypes of these repeatedly detected markers. Our results demonstrate the great potential of integrating the association analysis and marker-assisted breeding by utilizing the peanut mini-core collection.     

Diversity characterization and association analysis of agronomic traits in a Chinese peanut (Arachis hypogaea L.) mini-core collection

Association mapping is a powerful approach for exploring the molecular basis of phenotypic variations in plants.A peanut(Arachis hypogaea L.)mini-core collection in China comprising 298 accessions was genotyped using 109 simple sequence repeat(SSR)markers,which identified 554 SSR alleles and phenotyped for 15 agronomic traits in three different environments,exhibiting abundant genetic and phenotypic diversity within the panel.A model-based structure analysis assigned all accessions to three groups.Most of the accessions had the relative kinship of less than 0.05,indicating that there were no or weak relationships between accessions of the mini-core collection.For 15 agronomic traits in the peanut panel,generally the Q t K model exhibited the best performance to eliminate the false associated positives compared to the Q model and the general linear model-simple model.In total,89SSR alleles were identified to be associated with 15 agronomic traits of three environments by the Q t K model-based association analysis.Of these,eight alleles were repeatedly detected in two or three environments,and 15 alleles were commonly detected to be associated with multiple agronomic traits.Simple sequence repeat allelic effects confirmed significant differences between different genotypes of these repeatedly detected markers.Our results demonstrate the great potential of integrating the association analysis and marker-assisted breeding by utilizing the peanut mini-core collection.     

In vitro regeneration of peanut (Arachis hypogaea L.) through organogenesis: Effect of culture temperature and silver nitrate

Summary The effect of culture temperature on the morphogenetic response of Arachis hypogaea was studied. Cotyledons were cultivated on MS medium supplemented with 110 μM 6-benzyladenine. Leaf explants were cultivated in the presence of the same growth regulator at 22 μM. Cultures were incubated at temperatures of 25, 28, and 35±5° C. Both direct organogenesis from cotyledons and development of organogenic calluses from leaves showed optimal rates at 35±5° C. The highest frequency of elongation of buds into shoots from leaf-derived calluses occurred in the presence of 5 μM AgNO₃. At the best culture temperature, an average of 95% of shoots formed roots on growth-regulator-free MS medium. Plants were successfully transferred to soil, showing normal phenotypes.     

Utilizing peanut husk (Arachis hypogaea L.) in the manufacture of medium-density fiberboards

The main objective of this study was to investigate the potential of peanut husk (Arachis hypogaea L.) as a fiber–peanut mixture to produce fiberboards for general purposes. For panel production, the addition of peanut husk at various percentages to the wood fiber was the only variable. Panels produced utilizing peanut husk were compared to panels produced using 100% wood fiber. The chemical properties of peanut husk; holocellulose and lignin content, alcohol–benzene, hot and cold water, and dilute alkali (1% NaOH) solubility, were also determined. Results indicated that panels could be produced utilizing up to 30% peanut husk without affecting the usability of the panels. It was not possible to meet the minimum IB strength standards when peanut husk was added to the mixture. Higher additions resulted in panels having lower elastic and rupture moduli than the minimum requirements according to TS-EN standards.     

Fine mapping of qAHPS07 and functional studies of AhRUVBL2 controlling pod size in peanut (Arachis hypogaea L.)

Cultivated peanut (Arachis hypogaea L.) is an important oil and cash crop. Pod size is one of the major traits determining yield and commodity characteristic of peanut. Fine mapping of quantitative trait locus (QTL) and identification of candidate genes associated with pod size are essential for genetic improvement and molecular breeding of peanut varieties. In this study, a major QTL related to pod size, qAHPS07, was fine mapped to a 36.46 kb interval on chromosome A07 using F₂, recombinant inbred line (RIL) and secondary F₂ populations. qAHPS07 explained 38.6%, 23.35%, 37.48%, 25.94% of the phenotypic variation for single pod weight (SPW), pod length (PL), pod width (PW) and pod shell thickness (PST), respectively. Whole genome resequencing and gene expression analysis revealed that a RuvB-like 2 protein coding gene AhRUVBL2 was the most likely candidate for qAHPS07. Overexpression of AhRUVBL2 in Arabidopsis led to larger seeds and plants than the wild type. AhRUVBL2-silenced peanut seedlings represented small leaves and shorter main stems. Three haplotypes were identified according to three SNPs in the promoter of AhRUVBL2 among 119 peanut accessions. Among them, SPW, PW and PST of accessions carrying Hap_ATT represent 17.6%, 11.2% and 26.3% higher than those carrying Hap_GAC,respectively. In addition, a functional marker of AhRUVBL2 was developed. Taken together, our study identified a key functional gene of peanut pod size, which provides new insights into peanut pod size regulation mechanism and offers practicable markers for the genetic improvement of pod size-related traits in peanut breeding.     

Changes in auxin patterns in developing gynophores of the peanut plant (Arachis hypogaea L.)

The peanut plant (Arachis hypogaea L.) produces flowers aerially, but buries the recently fertilized ovules in the soil in order for the fruit and seeds to mature underground. The organ that carries the seeds into the soil is called the gynophore. The growth of the peanut gynophore is regulated primarily by indole-3-acetic acid (IAA). A monoclonal antibody raised against IAA was used to successfully detect and localize this growth substance in the tissues of developing peanut gynophores. Five different stages of development were analysed: (1) before fertilization; (2) after fertilization; (3) during downwards growth; (4) at soil penetration; and (5) at the early stages of fruit formation. While no auxin signal is visible in the unfertilized ovules and ovary region, an asymmetric signal is observed in the gynophore wall after fertilization. During downwards growth, the auxin signal is located in both the meristematic region and in the area encircling the seeds, as well as in the cortex and epidermis region of the elongation zone. Upon soil penetration, the auxin signal in the meristematic region disappears, and most of the signal is detected in the gynophore wall near the tip. At the early stages of peanut fruit development, auxin signal is found at the lowermost area of the bending fruit, which eventually causes the fruit to be positioned horizontally. The results of this study suggest that the possible source of auxin within the gynophore may be the area of the gynophore wall close to the tip.