Embryo Rescue in Wide Crosses in Arachis. 1. Culture of Ovules in Peg Tips of Arachis hypogaea

Interspecific hybridization in Arachis is restricted by earlyembryo abortion for many cross-combinations. Rescue of youngembryos in vitro within a week after fertilization is necessaryto recover these embryos before they abort. Peg tips, with theovule and embryo tissues, of A. hypogaea L. cv. ‘NC 6’,were cultured to compare ovule growth, callus production andpeg elongation. Tissues were collected 1, 2, 3 and 4 d afterself-pollination, after which peg meristems were removed fromhalf the pegs and cultured on five media combinations. One-day-oldpegs had significantly (P = 0.01) more ovule growth than oldertissues. Presence of the meristem had a greater inhibition toovule growth for 2- to 4-d pegs than for 1-d-old pegs. Significantlymore callus was produced on 4-d pegs than younger tissues, andkinetin had the greatest stimulatory effect on callus. Elongationof pegs with the meristem attached was observed most often inmedia with high sucrose levels. The observations indicate thatvery young ovules can be grown in vitro, and techniques maybe applicable to rescue of young embryonic tissues of Arachis. Ovule culture, interspecific hybridization, Arachis hypogaea, peanuts     

Early Reproductive Ontogeny in Interspecific Crosses of Arachis hypogaea and Section Arachis Species

Wild Arachis species have been recognized as sources of resistanceto pests and pathogens that infect A. hypogaea L. and causesubstantial yield losses. However, utilization of these geneticresources for crop improvement has been difficult. This studywas conducted to (a) understand the processes of early embryogrowth and development in four Arachis species, two A. hypogaeacultivars and their hybrids and (b) identify parental compatibilitiesin reciprocal crosses of A. hypogaea. The results indicatedthat delayed fertilization beyond 24 h, coupled with slow proembryogrowth, leads to embryo abortion in many interspecific crosses.For example, in female A. cardenasii crosses, lack of or delayedfertilization leads to failure to obtain hybrids. When A. batizocoiwas used as a female parent, delayed fertilization and the inabilityof quiescent proembryos to resume growth after soil penetrationcaused abortion. Embryos of A. hypogaea x A. glandulifera crossesdeveloped normally during the first 21 d after fertilization,but then aborted at a later time. In this study, A. hypogaeawas always a better female parent than the wild Arachis species.Increasing the number of pollinations per cross, using the cultivatedspecies as the female parent, utilizing different A. hypogaeavarieties, and embryo rescue techniques are suggested to improvethe probability of obtaining interspecific hybrids in Arachis.Copyright1995, 1999 Academic Press Peanut, interspecific hybrids, Arachis, wild species, incompatibilities     

Photomorphogenesis of the Gynophore, Pod and Embryo in Peanut, Arachis hypogaea L.

Darkened excized gynophores ceased to elongate after 8–10days in vitro and started to form a pod. Gynophore elongationwas inhibited to a greater extent in total darkness than underlow irradiance, while pod and embryo growth was stimulated indarkness only. Intact gynophores, enclosed in transparent vials containingglass beads, continued to elongate in both light and darkness.In light the elongating gynophores thickened as they penetratedbetween the glass beads, forming a seedless pod at the bottomof the vials. In the dark the elongating gynophores producedsmall pods in which the seeds had started to grow. Excized gynophores elongated in vitro under continuous whitelight at a rate similar to that of intact exposed gynophores.The rate of elongation in vitro, was lower under continuousblue or red-enriched light, than under white light, and wasfurther reduced under continuous far-red irradiation. Pods didnot form during any of the continuous irradiation treatmentsbut only after transfer to darkness, the largest pods formingafter continuous far-red irradiation. As little as 10 min daily exposure to red or far-red irradiancehad the same effect on gynophore elongation as continuous irradiation.Pods formed only when the daily periods of far-red irradiationwere 30 min or less. Reducing the daily exposures to 2 min decreasedthe time to onset of pod formation from 30 to 16 days. Far-redfollowing red irradiation was effective in inhibiting gynophoreelongation stimulated by red irradiation. Pod formation in red/far-redirradiation was only 50 per cent of that observed in far-redirradiation. The involvement of light in continual gynophoreelongation and in the concomitant inhibition of proembryo growthis discussed. Arachis hypogaea L., peanut, gynophore, photomorphogenesis, embryo development, pod development, proembryo     

Geotropic Responses and Pod Development in Gynophore Explants of Peanut (Arachis hypogaea L.) Cultured In Vitro

Peanut gynophore explants cultured in vitro on a defined mediumshow a positive geotropic response in both light and dark whenplanted either horizontally, or vertically with the tip pointingupwards. The growth following the initial curvature dependedon age of the gynophores and on the levels of growth substancesin the medium. In the dark and in presence of 0·01–0·1p.p.m. kinetin, naphthalene acetic acid at concentrations of0·1 p.p.m. and lower promoted gynophore elongation. Athigher concentrations elongation was promoted to a lesser extentin younger explants, caused enlargement of the ovary and formationof pods. Young explants generally elongated more than olderones and pod formation took place inside the medium, while inolder ones it took place above the medium. In the light, theinitial positive geotropic response was followed by elongationbut without any enlargement of the ovary. Decapitation of gynophores1·5–2·0 mm below their tip, removing theovary but leaving most of the intercalary meristem, had no effecton the geotropic response and elongation. The initial geotropicresponse and elongations of explants in vitro was not dependenton the presence of the ovary but on the meristem proximal toit. Changes in growth substances balance during gynophore developmentseem to affect geotropic response, elongation and pod formationin the peanut.     

Cultured cells of Arachis hypogaea susceptible to induction of stilbene synthase (resveratrol-forming)

Callus cultures were established from cotyledons of Arachis hypogaea. The cultured cells were shown to produce stilbene synthase (resveratrol-forming) if the cells were irradiated with 254 nm UV light. The lag period after which the stilbene synthase activity appeared was similar for cultured cells and intact cotyledons.     

光对不同叶龄花生叶片糖酵解代谢的影响

光对花生不同叶龄的磷酸丙糖、3-磷酸甘油醛脱氢酶、丙酮酸激酶和丙酮酸含量的影响不同。成长叶片光合速率大于暗呼吸,在光下,TP增高,细胞质G3PDh活性受抑制,PK活性和Pyr含量下降;DCMU处理则消除光的影响,TP下降,细胞质G3PDh和PK活性升高。花生成长叶在光下糖酵解(EMP)途径受抑制,抑制位点是G3PDh和PK。幼叶光合微弱,呼吸作用强,TP和Pyr及细胞质酶活均不受光、暗条件和DCMU处理影响,其EMP途径和在暗中同样速率运转。老叶光合和暗呼吸均较弱,TP和两种酶活不论在光和暗中均无差异,Pyr少,EMP途径运转也慢。     

Fertilization and Embryo Development in Wheat x Maize Crosses

The fertilization and embryo development in crosses of hexaploid wheat “Kangxuan 9” X maize “SS 7700” were studied. Of 180 florets fi,ced after pollination 34(18.9%) had embryo and endosperm, 46(25.6%) had only embryo and 12(6.7%) had only endosperm. Percentages of single or double fertilization were higher than that in control (“Chinese Spring” X maize). The hybrid embryos and endosperms obtained were karyotypically unstable and characterized by rapid elimination of the maize chromosomes to produce haploid wheat embryos. The potentials for wheat haploid production and transfer of DNA segments, including transposable elements, from maize to wheat is discussed.     

龙生型花生的遗传多样性

龙生型花生(Arachis hypogaea var.hirsute)是中国引进最早、种植最早的花生类型。经过不断地自然选择和人工选择,形成了许多各具特色的地方品种,并逐渐形成了次生中心。本实验对15份龙生型花生资源在植物学性状和品质性状及分子水平上进行了系统研究。结果表明:龙生型花生在植物学性状和品质性状上具有丰富的遗传多样性,在分子水平上差异也较大。变异系数在5.10~34.60之间,多样性指数在1.17~2.04之间,同时两者的变化趋势相反。基于植物学性状和品质性状上的聚类分析在阀值为6.5将龙生型花生分为两大类,一类主要包括广西的品种,另一类包括其它省份的品种;基于AFLP分析的聚类结果在阀值为0.39处分为5类,一类主要包括广西的品种,其它4类包括其它省份的品种。     

Comparison of Embryo Development in Wild and Cultivated Arachis Species

Embryo development following selfing was investigated in twowild diploid peanut species, Arachis batizocoi Krap. et Greg.(coll. K 9484) (2n = 20) and A. duranensis Krap. et Greg. nom.nud. (coll. K 7988) (2n = 20), and one cultivated tetraploidspecies, A. hypogaea L. NC-Ac 18000 (2n = 40). Rates of pegelongation and sequences of embryo development for each specieswere compared. Peg elongation rates were similar for the twowild species, but for A. hypogaea it was only one-third to one-halfthat of the diploid species. Embryos in A. hypogaea showed slightlymore rapid cell division than in the wild species. The observedvariation in reproductive development between the wild and cultivatedspecies indicate that different control mechanisms may governdevelopment in the different species and may be at least partiallyresponsible for failure to produce viable interspecific hybridsat various ploidy levels. The observations are also importantfor determining the time at which embryos of different speciesof Arachis will reach the appropriate stage of development forsuccessful culture on an artificial medium during embryo rescueprocedures. Peanut, Arachis hypogaea, wild species, embryo, peg     

Responses to Saturation Deficit in a Stand of Groundnut (Arachis hypogaea L.). 2. Growth and Development

Stands of groundnut were grown in four glasshouses with themaximum saturation deficit (D) of the air limited to 1.0, 2.0,2.5 or 3.0 kPa. The soil was near field capacity when plantsemerged and no water was applied thereafter. In a fifth glasshouse,a stand was grown at low D on soil irrigated to field capacityevery few days. Developmental processes such as timing of flowering, peggingand pod formation were unaffected by D, but the numbers of branches,flowers and pegs were reduced in the drier treatments. Measurementsduring the first 30 d showed that in the drier treatments leafgrowth was reduced, and the partitioning of dry matter intoroots was enhanced. In the unirrigated stands, dry matter production in shoots wasreduced by 40 per cent as the maximum D increased from 1.0 to3.0 kPa. Growth was affected through reductions both in leafarea (and therefore light interception) and in the productivityper unit of light intercepted. These responses to D and soilwater were linked to changes in bulk water potential of leaves. Productivity per unit of water transpired (q) decreased withincreasing D. The product of q and the mean daytime value ofthe difference in vapour pressure between leaf and air was moreconservative than q, and ranged from 3.1 to 5.6 g kPa kg^–1. Groundnut, Arachis hypogaea L., saturation deficit, growth, development, light interception, water use efficiency     

Diagnosis of iron deficiency in groundnut,Arachis hypogaea L.

Summary Investigations into iron deficiency have been hindered by the lack of a satisfactory diagnostic tissue test, which in turn results from the total iron content of plant tissue commonly being an unreliable index of the iron status. Our measurements of chlorotic and normal leaves of field grown groundnut (Arachis hypogaea L.) showed that total iron was unsatisfactory as the measure of iron status of plant tissue. It was found that iron status was better assessed from an estimate of the ferrous iron content of fresh leaf materials obtained by extraction with o-phenanthroline. Extractable iron content increased with leaf age. Chlorotic buds or the first fully opened leaf always contained less than 6μg extractable-Fe/g fresh tissue. Approved for publication as ICRISAT Journal Article No. 307.     

Characterization of isoperoxidase activity in Arachis hypogaea cultivars

Isoperoxidases (both anodic and cathodic) from individual seeds of several peanut cultivars (Arachis hypogaea), after ammonium sulfate precipitation of the major storage protein, arachin, from the extracts were characterized by polyacrylamide gel electrophoresis with respect to seed development, maturity and germination and the geographic areas where grown. All cultivars had a major cathodic isozyme near the origin of the gels. Much anodic intra- and inter-varietal isozyme polymorphism was observed in mature seeds collected from different geographic areas. These polymorphic isozymes were consistently present during the earlier stages of seed development (the activities decreased quantitatively and became variable during the later stages of maturation), and most were observed in each peanut upon germination, the latter showing quantitative increases in activity.     

Genome size variation in Arachis hypogaea and A. monticola re-evaluated.

Genome size variation within species is a frequently reported, but still a controversial problem. In the present study, we re-evaluated recently published Feulgen densitometric data on genome size and its infraspecific variation in Arachis hypogaea, and also conducted measurements in one accession of its wild relative A. monticola. The methods applied were propidium iodide flow cytometry and Feulgen densitometry using Pisum sativum as an internal standard. The 2C DNA contents previously published cannot be confirmed, but values obtained in this study are about half as large. Additionally, we could not reproduce the previously reported 1.15-fold variation within A. hypogaea; our data indicate genome size stability between respective accessions of this species. Based on 8.84 pg (2C) for Pisum sativum the DNA amounts (2C) were: 5.914 pg in A. hypogaea, and 5.979 pg in A. monticola.     

The Morphology and Anatomy of Ovule and Fruit Development in Arachis hypogaea L

The vascular system of the monocarpellary gynoecium with tenwell differentiated traces and a few cross links probably representsa precocious development of the post-fertilization vasculatureof the fruit wall. The restriction of the two integuments ofthe ovule to the micropylar half, and the endothecial natureof the chalazal cells adjoining the embryo sac appear to indicatea pathway of derivation of the unitegmic tenumucellate ovulefrom the bitegmic crassinucellate one. During double fertilization,a dark staining refractive body appears in the nucleolus ofthe egg as well as the fusion product of the polar nuclei. The peg that carries the ovary into the soil after fertilizationgrows by the activity of a rib meristem at the basal solid partof the gynoecium. During sub-soil fruit development, the ovarywall develops a prominent spongy inner zone which finally disappears,and a peripheral zone that forms the mature fruit wall. Theabinitio nuclear endosperm is much reduced and degenerates afterproducing a few cell layers in the chalazal half alone. Seeddevelopment is pachychalazal. The main vascular supply of theseed branches at the chalaza into eight to ten strands in theseed coat. All seeds that have a vascular ramification in theseed coat are probably pachychalazal. In the variety Valencia, diminutive fruits with viable seedmay develop aerially from pegs that fail to grow long enoughto reach the soil from the higher nodes. Arachis hypogaea L., groundnut, fruit development, seed development, carpel vasculature, seed vasculature, pachychalaza     

Role of carbohydrate moieties in peanut (Arachis hypogaea) peroxidases.

The activities of a cationic (C.PRX) and an anionic peroxidase isolated from peanut (Arachis hypogaea)-cell suspension culture were drastically reduced when they were deglycosylated with glycopeptidase F or oxidized by 10 mM-periodate. In contrast with the controls, the deglycosylated or the oxidized peroxidases were much more susceptible to proteolytic degradation. In radiolabelling experiments with [35S]methionine, the non-glycosylated C.PRX was synthesized in the tunicamycin-treated cultures and secreted into the medium. Examination of the C.PRX polypeptides by SDS/polyacrylamide-gel electrophoresis followed by fluorography showed that the non-glycosylated form had an Mr of approx. 31,000, which is about 78% of that of the glycosylated form. Our results suggest that carbohydrates may not be essential for peroxidase secretion, but that stabilization of the peroxidase molecules and acquisition by these isoenzymes of a catalytically active conformation is linked directly or indirectly to glycosylation.     

Utilization of wild relatives in genetic improvement of Arachis hypogaea L.

Summary The chromosome complements of 12 taxa in section Arachis were karyotypically and meiotically analysed. In taxa with 2n=20 the arm ratio of the respective pair of chromosomes was taken as an independent quantitative character and statistically analysed by Mahalanobis D². Two clusters were formed, one represented solely by A. batizocoi and the other consisting of the remaining 11 taxa. This grouping was confirmed by canonical analysis. In the larger group of species, A villosa and A. correntina were closely related karyotypically and on D² distance, while A. cardenasii forms a distinct subgroup. A. cardenasii lacks the short A chromosome recorded in other species of this group, and A. batizocoi is no longer the only species to have a pair of chromosomes with a secondary constriction. The taxa with 2n=40, A. monticola and A. hypogaea, are karyotypically very similar, though there is a difference in the number of chromosome pairs with a secondary constriction. On the basis of karyomorphological affinity, especially in relation to marker chromosomes, A. cardenasii is probably one of the ancestors of the tetraploid species studied.Approved as ICRISAT Journal Article No. 169 and released for publication     

Development of Photosynthetic Structure and Function during Light-Induced Greening in Photomixotrophic Cells of Arachis hypogaea L.

The synthesis of chlorophyll and development of photochemicalactivities were complete within 70–80 h in greening leaves,whereas these processes continued for 8 days with an initiallag of 8 h for pigment synthesis in greening Arachis hypogaeaL cells. The activity of photosystem I in cultured Arachis cellswas detected earlier (24–36 h after illumination) thanthat of photosystem II (42–54 h after illumination) andthe development of the latter coincided with the synthesis ofa 46,000 dalton polypeptide of the thylakoid membranes. Experimentalstudies with cultured cells have the advantage in that the temporalsequence of the assembly of membrane components and associatedfunctions are determined easily because of longer developmentalperiod of chloroplast. (Received January 29, 1982; Accepted April 13, 1983)     

Effects of Growth Regulators on Incompatible Crosses in the Genus Arachis L.

Wild species in the genus A rachis L. have several desirablecharacters and are therefore of great interest for use in thegenetic improvement of A. hypogaea L. but only a few closelyrelated species have been used. There are many others whichcould not be used because they are incompatible with the cultivatedspecies. This paper reports techniques to produce hybrids fromincompatible crosses.     

Absence of Root Hairs in Non-Nodulating Groundnut, Arachis hypogaea L.

Root hairs observed at the site of lateral root emergence innodulating groundnut cultivars were found to be absent in non-nodulatinggroundnut lines. In a segregating F₂-population of the crossNC 17 x PI 259747 a strong association was observed betweenthe presence of root hairs and nodulation, and the absence ofroot hairs and non-nodulation. Key words: Root hairs, Arachis hypogaea, Non-nodulation     

Some Observations on Infection of Arachis hypogaea L. by Rhizobium

The infection process in Arachis hypogaea by rhizobia differsfrom that normally found in Trifolium spp. in that no infectionthreads are formed. The root hairs, which are long (up to 4mm), septate, and often with large basal cells, occur only atthe sites of emerging lateral roots. Infection occurs only wherethe root hairs have large basal cells. Rhizobia cause curlingand deformation of the root hairs (as in Trifolium spp.) butenter the root at the junction of the root hair and the epidermaland cortical cells. The bacteria are distributed intercellularlyvia the middle lamellae and enter the cortical cells throughthe structurally altered cell wall, often close to the hostcell nucleus. The root hairs and large basal cells become infectedin the same way. Within the cortical cells of the emerging lateralroot the rhizobia multiply rapidly and the invaded cells dividerepeatedly to form the nodule tissue. Bacteriod formation occursonly when the host cell ceases to divide.