花生体细胞胚胎发生的组织细胞学研究

成熟花生胚轴在MS 40mg/\L2,4-D 0.5mg/L KT培养基中可直接诱导产生体细胞胚，降低2，4-D的浓度，将诱导产生的体细胞胚组织块继代2-3次可诱导产生出米黄色颗粒状的胚性愈伤组织，继而产生体细胞胚。花生体细胞胚直接诱导起源于表皮或近表皮的细胞，其形成经历了跟合子胚相似的过程。花生胚性愈伤组织的细胞具有细胞小、胞质浓、核大、核仁明显，排列紧密的特点。经愈伤组织形成的体细胞胚以单细胞内起源方式发生。首先由体细胞胚单个原始细胞分裂形成2-细胞原胚，2-细胞原胚继续分裂成3-细胞原胚或4-细胞原胚，3-细胞原胚或4-细胞原胚继续分裂形成多细胞原胚，进一步发育形成一个完整的体细胞胚。     

Reduced trigonelline accumulation due to rhizobial activity improves grain yield in peanut (Arachis hypogaea L.)

Abstract

Crop improvement for drought tolerance is critical for the future of crop production. The objectives were to examine the relationship between trigonelline (TRG) accumulation and yield traits in 10 peanut (Arachis hypogaea L.) genotypes inoculated with two commonly used nitrogen-fixing Rhizobium spp., and to evaluate a role of TRG on growth traits. TRG increased as a defensive metabolite in response to water deficit, but resulted in the reduction of the number of nodules and yield. Symbiotic rhizobial activity helped plants to improve yield particularly in a fully irrigated field rather than under reduced irrigation. TRG concentrations in genotypes (7 out of 10) increased under reduced irrigation as compared with those under full irrigation in two years. Mean number of nodules at maturity (120 days after planting) across genotypes under reduced irrigation were 89 in the control, 111 in Lift, and 161 in Histick treatments, among which Histick was significant (p < 0.05) for nodulation. Mean pod yields in the control, Histick, and Lift treatments were 1.69, 2.34 and 1.87 Mg ha^-1, respectively, under reduced irrigation. Under full irrigation, pod yields were 3.35 in the control, 4.50 in Histick and 3.41 Mg ha^-1 in Lift treatments, but were more significantly improved as treated with Histick than other treatments. Genotype ICGS-76 produced the highest pod yield (5.13 Mg ha^-1) as treated with Histick. All genotypes treated with Histick and Lift biosynthesized less TRG (decreased from 5.8 to 65.3% relative to the control) but produced larger numbers of pods (increased from 9.2 to 80.4% relative to the control), which resulted in substantially higher pod yields.     

Silicon Effect on Nutrient Acquisition of Peanut (Arachis hypogaea L.) Under Aluminum Stress

Aluminum (Al) toxicity represents one of the main yield-limiting factors for crops in acid soils. Silicon (Si) is known to increase tolerance in higher plants. This study was conducted to determine whether treatment with Si could improve nutrient uptake by peanut under Al stress. Peanut (Arachis hypogaea L. cv Zhonghua 4) was raised with or without Si (1.5 mM) in the growth chamber under 0 and toxic Al (0.3 mM) levels. Aluminum stress significantly decreased the root- and total-dry weight by 52.4% and 32.0%, respectively. The content of nitrogen (N), phosphorus (P), potassium (K), calcium (Ca), and magnesium (Mg) was significantly decreased, but that of Al increased markedly in shoots and roots after Al exposure at seedling, flower-needle, and pod-setting stage. Silicon alleviates Al toxicity in peanut plants in relation to Al distribution and allocation of tissue P, K, Ca, and Mg by favoring the partitioning of dry mass to roots.     

Evaluation of genetic relationships among botanical varieties of cultivated peanut (Arachis hypogaea L.) using AFLP markers

Forty-four accessions of cultivated peanut (Arachis hypogaea L.) representing sixbotanical varieties of two subspecies along with three accessions ofthe wild relative A. monticola Krapov et Rigoni were evaluated for their genetic relationships using theAFLP marker technology. Fifteen AFLP primer pairs (EcoRI/MseI) generated 28distinct polymorphic markers that were employed to develop uniqueprofiles of all accessions and to construct a phenogram. The resultsshowed that the botanical varieties aequatoriana and peruviana werecloser to subspecies hypogaea than subspeciesfastigiata Waldr. to which they belong, and the wildA. monticola was notdistinct from the cultivated A.hypogaea. Although the extent of geneticdiversity in peanut is low compared to many other crops, our studiesshow that by employing the AFLP approach, sufficient DNA variationcan be detected in the cultivated peanut germplasm to conductevolutionary studies.     

花生栽培种InDel有效标记筛选与评估

为获得花生栽培种中有效的InDel标记,本研究基于花生简化基因组检测到的InDel信息,经PCR扩增以及电泳检测验证引物的有效性与多态性,利用生物统计学软件检测InDel标记的多态性水平。结果表明,在花生基因组上设计的39对引物在覆盖4个植物学类型的21份花生中均可获得扩增产物,其中14对引物具有多态性,为总引物数的35.9%,有3对引物位于基因编码区;Shannon指数为0.191 4～1.295 1,均值为0.579 4;多态性信息量(PIC)为0.090 7～0.674 6,均值为0.349 6。本研究筛选获得的InDeI标记可为花生栽培种遗传图谱构建和分子标记辅助育种提供有效的遗传标记。     

Non-rhizobial peanut nodule bacteria promote maize (Zea mays L.) and peanut (Arachis hypogaea L.) growth in a simulated crop rotation system

The term “Plant Growth Promoting Bacteria” or PGPB designates a diverse group of prokaryotic microorganisms that can increase plant growth by diverse mechanisms. Some PGPB are capable of colonizing root inner tissues and constitute endophytic populations. Incorporation of these microorganisms into agricultural practices may constitute a valid alternative to increase crop productivity in a sustainable and environmentally friendly production scheme, reducing the application of agrochemicals. In a previous work, we described the characterization of bacteria belonging to Pseudomonas, Enterobacter and Klebsiella obtained from surface sterilized peanut nodules. In addition, we showed that some of these isolates were able to promote several peanut growth and symbiotic parameters. Bounded to the results from this particular study, and considering their potential ability to interact with different plant species, in this work we assessed the effects of their inoculation in maize (Zea mays L.) under controlled conditions. Furthermore, we analyzed growth promotion in a simulated peanut–maize crop rotation system. Finally, we determined the plant growth promoting (PGP) properties present in the isolates. Results indicated that all bacteria are able to significantly promote maize and peanut growth, and that they also displayed plant growth promotion activity in maize growing in a peanut–maize crop rotation sequence.     

花生品种间杂种F1代的SSR标记分析

以花育22号和Sunoleic95R为亲本配置杂交组合,获得杂种F1,利用SSR引物对亲本和杂种F1各单株进行扩增,分析其带型,辨别真伪杂种,并对亲本和杂种F1各单株的农艺性状、品质进行调查测定。结果表明：5对引物PM50 、PM179、 PM15、PM384和PM348对亲本、杂种F1各单株的分析结果是一致的,15个杂种F1单株,8株表现为母本带型,是伪杂种;7株表现为杂合带型或父本带型,即真杂种。但5对引物的带型表现存在明显差异,PM50 、PM179、 PM15、PM384表现为双亲互补带型, PM348表现为偏父本型。农艺性状和品质性状分析表明真杂种与伪杂种的籽仁重、侧枝长、亚油酸含量、油酸含量差异大,主茎高、蛋白质含量、脂肪含量差异较小。证实利用SSR标记辨别栽培种花生真伪杂种是可行的。     

Inheritance of iron deficiency chlorosis resistance in groundnut (Arachis hypogaea L.)

Iron-deficiency chlorosis (IDC) is an important abiotic constraint affecting the growth and yield of groundnut in calcareous and alkaline soils worldwide. The present study investigated the inheritance of IDC resistance among four straight crosses of groundnut involving four IDC susceptible cultivars as females and a common IDC resistant male parent. The F₁'s of all the four crosses were resistant to IDC indicating the dominant nature of IDC resistance. The F₂'s of all the four crosses showed a good fit to the ratio of 15 (IDC resistant): 1 (IDC susceptible) and their behavior among the F₃'s was as per the expected ratio of 7:4:4:1. The IDC resistance in groundnut is under the control of duplicate dominant genes wherein, the presence of a dominant allele at either of the loci results in IDC resistance, while duplicate recessive results in IDC susceptibility. This information would facilitate development of IDC resistant cultivars of groundnut.     

Sodium azide induced high-oleic peanut (Arachis hypogaea L.) mutant of Virginia type

Less prone to oxidation than its conventional counterpart, high-oleic peanut (Arachis hypogaea L.) containing?≥?72% oleate and?<?8% linoleate is preferred by processors, seed sellers and consumers. Development of high-oleic peanut cultivars with suitable pod/seed size and shape may satisfy the need from food processors manufacturing whole kernel or halve products. In this study, a high-oleic Virginia type peanut mutant was identified through screening of a 15 mmol/L sodium azide mutagenized M3 population with near infrared spectroscopy. Sequencing of the mutated and wild type FAD2A/FAD2B genes detected 2 point mutations. The G448A mutation in FAD2A was the same as in previous reports, causing an amino acid change of D150N. G558A in FAD2B was a novel mutation, resulting in a stop codon and premature termination of protein synthesis. 16 promising lines with acceptable productivity and pod/seed characters have been tentatively bred, which will be evaluated further in yield tests with replications after seed increase.

     

The genome donors of the groundnut/peanut (Arachis hypogaea L.) revisited

Arachis hypogaea, the cultivated groundnut is a tetraploid with an AABB genomic constitution. The available literature on the origin of groundnut reveals that there is general agreement that the cultivated groundnut has evolved from the wild tetraploid species A. monticola, with which it crosses freely to produce fertile hybrids. However, the issue of actual diploid ancestors of A. monticola is still unresolved. Both cytogenetic and molecular evidences support A. duranensis being the most probable progenitor and donor of the A genome to A. hypogaea. For the B genome, the cytogenetic evidence suggests A. batizocoi to be the most probable progenitor, but the RFLP banding pattern indicates that A. batizocoi is more distantly related to A. hypogaea than other species of section Arachis. RFLP banding pattern indicates A. ipaensis to be one of the closest species to A. hypogaea and the possible donor of the B genome. The present article critically analyzes the available data, which suggests that until an amphidiploid is produced synthetically between A. duranensis × A. ipaensis and crossed successfully with A. hypogaea to produce a fertile hybrid, this issue would remain unresolved. A. batizocoi would remain the most probable donor of the B genome because of its directly demonstrable cytogenetic affinity.     

Diagnosis of zinc deficiency in peanut (Arachis hypogaea L.) by plant analysis

Abstract

Critical values of zinc (Zn) concentration in young leaves Here established for the diagnosis of Zn deficiency in peanut by examining the relationship of Zn concentration in leaves to shoot dry matter (DM) at two growth stages of plants grown in pots of Zn deficient sand at seven levels of Zn supply (0, 67, 133, 200 267, 533, and 1067 μg Zn/kg soil). Zinc deficient peanut accumulated reddish pigments in stems, petioles and leaf veins in addition to the more common symptoms of Zn deficiency in plants. Zinc concentrations increased with increasing Zn supply in the blades of the youngest fully expanded leaf (YFEL) and in the blades of the leaves immediately older (YFEL+1) and younger (YFEL‐1) than it: they also increased with increasing Zn supply in the petioles of the YFEL+1 and YFEL and in the basal stem internode but their Zn concentrations Here always much lower than those in the blades. Critical Zn concentrations in the blades of the YFEL and YFEL+1 Here 8–10 mg Zn/kg DM at early pegging and mid pod filling: values for YFEL‐1 were similar but more variable. The blade of the YFEL is recommended for diagnosis of Zn deficiency in peanut and 8–10 mg Zn/kg DM as its critical value.     

Soil fertility effects on growth and nutrient uptake of rhizoma peanut

Expansion of forage production acreage for rhizoma peanut (RP ‐ Arachis glabrata Benth.), a persistent perennial, warm‐season forage legume adapted to the Gulf Coast region of the southeastern United States, is limited primarily by establishment rate, often taking up to three years. The objective was to determine the effects of two divergent soil environments on the dry matter production and nutrient uptake of RP in the first year following establishment. Five Arachis genotypes, ‘Arbrook’, ‘Florigraze’, and experimental lines 423, 575, and 616, were established during March 1995 at Clinton and Olive Branch, LA. Soils were a Dexter silt loam (Ultic Hapludalf) and a Loring silt loam (Typic Fragiudalf, fine‐silty, mixed, thermic) with mean pH values of 6.8 and 4.7, respectively. Soils also differed for extractable potassium (K), sodium (Na), calcium (Ca), aluminum (Al), sulfur (S), copper (Cu), iron (Fe), manganese (Mn), and organic matter (OM). Tissue samples were taken at three week intervals in 1996 and yield and quality were evaluated at six week intervals. Locations differed significantly for dry matter production, crude protein (CP), neutral detergent fiber (NDF) and in vitro true digestibility (IVTD). Entries also differed within locations for yield, CP, NDF, acid detergent fiber (ADF), and IVTD. Tissue nitrogen (N), K, Ca, S, Mn, boron (B), and zinc (Zn) were significantly different between locations, but phosphorous (P), magnesium (Mg), Fe, Cu, and Al were not different. Yield and forage quality were both inversely correlated with pH and, with the exception of N, none of the other nutrients were low enough to account for the differences observed in growth between the two locations.     

硅、钙对花生新品种郑农花9号的生育及产量的影响

以花生品种郑农花9号为材料进行硅、钙基肥试验,基施硅钙镁肥、高效硅肥、钙肥,探讨硅、钙元素对花生生长及产量的影响。结果表明:3种肥料均可使花生茎秆粗壮、生长旺盛、抗逆性增强,病害发生率降低;硅钙镁肥使花生出苗期提前2 d,出苗率、单株结果数优于钙肥和高效硅肥。所有处理荚果产量增产41.7～717.0kg/hm2,增产幅度0.85%～14.7%。钙肥对花生产量构成因素的影响最大,第一对侧枝的平均长度最长,单株饱果数平均11.15个,平均百果重273.25 g、百仁重117.25 g,出米率和单株饱果数都是最高,钙肥的平均单产为5 386.01 kg,其3个处理增产极显著,硅钙镁肥的平均单产为5 260.95 kg,其两个处理增产极显著,高效硅肥的平均单产为5 190.08 kg。含钙的肥料组合较之无钙的肥料对产量及其构成因素的影响显著,更为可靠。     

Morphological and reproductive characterization of nascent allotetraploids cross-compatible with cultivated peanut (Arachis hypogaea L.)

Genetic Resources and Crop Evolution - Peanut improvement is limited by a narrow genetic base. However, this obstacle can be circumvented by incorporating phenotypic variability from wild, diploid...     

Competition among strains of Bradyrhizobium and vesicular-arbuscular mycorrhizae for groundnut (Arachis hypogaea L.) root infection and their effect on plant growth and yield

Summary Strains of Bradyrhizobium influenced root colonization by a species of vesicular-arbuscular mycorrhizae (VAM), and species of VAM influenced root nodulation by strains of Bradyrhizobium in pot experiments. In a field experiment, the effects of VAM on competition amongst inoculated bradyrhizobia were less evident, but inoculation with Bradyrhizobium strains increased root colonization by VAM. Certain VAM/Bradyrhizobium inoculum strain combinations produced higher nodule numbers. Plants grown without Bradyrhizobium and VAM, but supplied with ammonium nitrate (300 g ml^–1) and potassium phosphate (16 g ml^–1), produced higher dry-matter yields than those inoculated with both symbionts in the pot experiment. Inoculation with either symbiont in the field did not result in higher pod and haulm yields at harvest.ICRISAT Journal Article No. 886     

花生种子特异启动子AHSSP1的克隆及功能分析

为丰富花生种子特异启动子资源,本研究利用PCR技术在花生基因组中克隆了种子贮藏蛋白基因PSC32的启动子AHSSP1,利用半定量RT-PCR检测了PSC32基因表达模式,借助NewPLACE在线分析了AHSSP1序列中存在的顺式作用元件,并构建了AHSSP1驱动GUS报告基因的表达载体,经农杆菌转化获得转基因拟南芥,经GUS组织化学染色鉴定了该启动子的功能。结果表明,PSC32基因957 bp长的启动子AHSSP1序列具备种子特异表达启动子特有的3个RY REPEAT元件。半定量RT-PCR分析发现,PSC32基因在花生成熟种子中表达,而在饱果成熟期根、茎、叶片、花、入土前的果针、成熟种子的果壳中均不表达。GUS组织化学染色发现,转基因拟南芥成熟种子以及萌发种子的子叶、下胚轴和胚根均能够被染上蓝色;长出真叶后,子叶和下胚轴仍能被染色,而根和真叶不能被染上蓝色;成年期转基因拟南芥的叶片也不能被染上蓝色。而野生型拟南芥整个生长时期均不能被染上蓝色。以上现象说明AHSSP1是一个种子特异启动子。本研究丰富了花生种子特异启动子的资源,对花生籽仁品质改良或以花生籽仁作为"生物反应器"的研究具有重...     

花生C4H和ANR基因的克隆与表达研究

通过构建花生未成熟种子cDNA文库,结合大规模EST测序,首次从花生中克隆了原花青素代谢途径上的2个关键酶［肉桂酸-4-羟化酶(C4H)和花青素还原酶(ANR)］基因的全长编码序列。序列分析表明,与其他植物的C4H相比,花生C4H整个编码区高度保守,开放阅读框长度1518bp,编码蛋白长度505个氨基酸,预测的蛋白分子量为57.9kDa,等电点为9.04。花生中ANR开放阅读框长度966bp,编码蛋白长度321个氨基酸,预测的蛋白分子量为35kDa,等电点为8.31。亚细胞定位预测分析表明,C4H定位在线粒体中,ANR则定位在胞外。半定量RT-PCR结果表明,C4H在花生根、茎、叶、花、果针、种子中均有表达,但在茎、叶中表达相对较低,而原花青素合成途径中的关键酶ANR在各个组织中均有表达,其中在果针中表达最强。     

Biosynthesis enhancement and antioxidant and anti-inflammatory activities of peanut (Arachis hypogaea L.) arachidin-1, arachidin-3, and isopentadienylresveratrol

Peanut is a potent plant to be induced to synthesize bioactive stilbenoids. Bioactivities of those stilbenoids except resveratrol have been meagerly investigated. When peanut kernels (Tainan 14, a Spanish cultivar) were imbibed, incubated 3 days for germination, sliced, incubated with artificial aeration, periodically sampled, lyophilized, extracted with methanol, and subjected to reverse-phase HPLC analysis, four major fractionations were detected and identified as trans-resveratrol (Res), trans-arachidin-1 (Ara-1), trans-arachidin-3 (Ara-3), and trans-isopentadienylresveratrol (IPD). During incubation of the peanut slices, contents of Res, Ara-1, and Ara-3 increased tremendously from initially trace or not detectable amounts up to 147.3, 495.7, and 2414.8 microg/g, corresponding to 20, 16, and 24 h of incubation, while IPD contents continued to increase up to 28 h (4474.4 microg/g). When the four stilbenoids and butylated hydroxytoluene (BHT) were subjected to antioxidant characterization by various measures, all have exhibited varied potencies of antioxidant activity. In particular, retardation of absorbance increase at 234 nm as formation of the conjugated diene hydroperoxides in a real pork oil system stored at 60 degrees C, supplement of Ara-1 at 100 microM has shown equivalent or even greater activity than did BHT. When the media were supplemented with Res, Ara-1, Ara-3, and IPD at 15 microM for cultivation of mouse macrophage RAW 264.7 cells activated by lipopolysaccharide (LPS), the LPS-induced extracellular production of prostaglandin E2 (PGE2) and nitric oxide (NO) was significantly inhibited by Ara-1 (p < 0.001), Res (p < 0.001), Ara-3 (p < 0.01), and IPD (p < 0.01). It is noteworthy and of merit that all test stilbenoids have exhibited potent antioxidant and anti-inflammatory activities and varied as affected by number of hydroxyl groups and isopentenyl or isopentadienyl moiety. Keywords: Arachis hypogaea L.; peanut; groundnut; resveratrol; stilbenoids; arachidin; antioxidant; anti-inflammation.