Construction of Genetic Linkage Map Based on SSR Markers in Peanut(Arachis hypogaea L.)

Molecular genetic maps of crop species can be used in a variety of ways in breeding and genomic research such as identification and mapping of genes and quantitative trait loci （QTLs） for morphological, physiological and economic traits of crop species. However, a comprehensive genetic linkage map for cultivated peanut has not yet been developed due to the extremely low frequency of DNA polymorphism in cultivated peanut. In this study, 142 recombinant inbred lines （RILs） derived from a cross between Yueyou 13 and Zhenzhuhei were used as mapping population in peanut （Arachis hypogaea L.）. A total 652 pairs of genomic-SSR primer and 392 pairs of EST-SSR primer were used to detect the polymorphisms between the two parents. 141 SSR primer pairs, 127 genomic-SSR and 14 EST-SSR ones, which can be used to detect polymorphisms between the two parents, were selected to analyze the RILs population. Thus, a linkage genetic map which consists of 131 SSR loci in 20 linkage groups, with a coverage of 679 cM and an average of 6.12 cM of inter-maker distance was constructed. The putative functions of 12 EST-SSR markers located on the map were analyzed. Eleven showed homology to gene sequences deposited in GenBank. This is the first report of construction of a comprehensive genetic map with SSR markers in peanut （Arachis hypogaea L.）. The map presented here will provide a genetic framework for mapping the qualitative and quantitative trait in peanut.

Drought-induced responses of organic osmolytes and proline metabolism during pre-flowering stage in leaves of peanut (Arachis hypogaea L.)

Peanut （Arachis hypogaea L.）, an improtant oil crop, usually encounters drought stress in the process of growth and development, especially at pre-flowering stage. In order to gain insight into the drought tolerance potentials based on osmolyte accumulation and metabolism of proline aspects of peanut, pot experiments were conducted with a split-plot design in Tai＇an, Shangdong Province, China in 2013 and 2014. Pre-flowering drought （PFD） stress and optinum irrigation （control, CK） were served as the main plots and the two peanut cultivars Shanhua 11 and Hua 17 served as sub-plots. Shanhua 11 was drought-tolerant cultivar and Hua 17 was drought-sensitive. The content of soluble sugars, soluble protein, free proline and other free amino acids, the activities of enzymes involved in proline metabolism, and malondialdehyde （MDA） content and ion leakage were all investigated in the two cultivars at pre-flowering stage. Results showed that PFD stress significantly increased the levels of soluble protein, free proline and free amino acid, and increased Al-pyrroline-5-car- boxylate synthetase （P-5-CS, EC 2.7.2.11） activity in the leaves of drought-tolerant and drought-sensitive cultivars. The activity of proline dehydrogenase （proDH） （EC 1.5.99.8） decreased under PFD stress in both cultivars. The leaves of the tolerant cultivar maintained higher increments of osmolyte levels, lower increments of MDA content and ion leakage, as well as a higher increased proportion of P-5-CS activity and higher inhibited proportion of proDH activity under water stress compared with the drought-sensitive cultivar. The study suggests that proline accumulation in peanut leaves under PFD can be explained by the higher enhanced activities of P-5-CS and higher inhibition of proDH. The results will provide useful information for genetic improvement of peanut under drought tolerance.

Functional Analysis of the Phosphoenolpyruvate Carboxylase on the Lipid Accumulation of Peanut(Arachis hypogaea L.) Seeds

Phosphoenolpyruvate carboxylase（PEPC;EC 4.1.1.31） catalyses phosphoenolpyruvate（PEP） to yield oxaloacetate,which is involved in protein biosynthesis.Pyruvate kinase（PK;EC 2.7.1.40） catalyzes PEP to yield pyruvate,which is involved in fatty acid synthesis.In this study,five PEPC genes（AhPEPC1,AhPEPC2,AhPEPC3,AhPEPC4,and AhPEPC5） from peanut have been cloned.Using a quantitative real-time RT-PCR approach,the expression pattern of each gene was monitored during the seed development of four peanut varieties（E11,Hebeigaoyou,Naihan 1,and Huayu 26）.It was found that these five genes shared similar expression behaviors over the developmental stages of E11 with high expression levels at 30 and 40 d after pegging（DAP）;whereas these five genes showed irregular expression patterns during the seed development of Hebeigaoyou.In Naihan 1 and Huayu 26,the expression levels of the five genes remained relatively high in the first stage.The PEPC activity was monitored during the seed development of four peanut varieties and seed oil content was also characterized during whole period of seed development.The PEPC activity followed the oil accumulation pattern during the early stages of development but they showed a significantly negative correlation thereafter.These results suggested that PEPC may play an important role in lipid accumulation during the seed development of four peanut varieties tested.

Identification of main effect and epistatic QTLs controlling initial flowering date in cultivated peanut(Arachis hypogaea L.)

Initial flowering date(IFD)is closely related to mature period of peanut pods.In present study,a population of recombinant inbred lines(RIL)derived from the cross between Silihong(female parent)and Jinonghei 3(male parent)was used to map QTLs associated with IFD.The RIL population and its two parental cultivars were planted in two locations of Hebei Province,China from 2015 to 2018(eight environments).Based on a high-density genetic linkage map(including 2996 SNP and 330 SSR markers)previously constructed in our laboratory,QTLs were analyzed using phenotypic data and the best linear unbiased prediction(BLUP)value of initial flowering date by inclusive composite interval mapping(ICIM)method.Interaction effects between every two QTLs and between individual QTL and environment were also analyzed.In cultivated peanut,IFD was affected by genotypic factor and environments simultaneously,and its broad sense heritability(h2)was estimated as 86.8%。Using the IFD phenotypic data from the eight environments,a total of 19 QTLs for IFD were detected,and the phenotypic variation explained(PVE)by each QTL ranged from 1.15 to 21.82%.Especially,five of them were also detected by the BLUP value of IFD.In addition,12 additive QTLs and 35 pairs of epistatic QTLs(62 loci involved)were identifed by the joint analysis of IFD across eight environments.Three QTLs(qIFDB04.1,qIFDB07.1 and qIFDB08.1)located on chromosome B04,B07 and B08 were identified as main-effect QTL for IFD,which had the most potential to be used in peanut breeding.This study would be helpful for the early-maturity and adaptability breeding in cultivated peanut.

A New Coumestan from Arachis hypogaea L.

A new coumestan, 3, 9-dihydroxy-4, 8-dimethoxycoumestan, was isolated from Arachis hypogaea L. together with two known compounds: 3, 9-dihydroxy-4-methoxycoumestan and 3, 9-dihydroxy-8-methoxycoumestan. The structure was established by spectroscopic methods.

Gomparative analysis of DGAT3（diacylglycerol acyltraiisferase 3） gene from different peanut （Arachis hypogaea L.） varieties

Cultivated peanut is one of the primary sources of vegetable oil and protein in developing countries. DG〉A73 family in peanut cotyledons has no membrane-bound regions suggesting that cytosol is one of the sites for triacylglycerol （TAG） biosynthesis in oilseeds. According to functional annotation and classification of 5 cDNA libraries, 12 unigenes were found with relation of peanut DGAT3 in different organs. Three clones of unigenes, OCP- contig168t OCPcontig12101-3 and OCPcontigl2388-1 were selected for sequencing. Full length sequence of DGAT3 was obtained, showing over 98% sequence similarity with peanut DGAT3 gene AY875644 or EU183333. Upon cluster analysis, DGAT3 of 40 culti- vars were divided into 3 types, namely AhDGATS-1, AhDGAT3-2 and AhDGAT3-3. Coding regions are 1023, 1038 and 1026 base pairs which encoded proteins with 340-, 345- and 341-amino acids, respectively. DGAT3-3 might be a novel gene type among the DGAT3 family which provides great help for studying DGAT3 gene evolution in peanut.

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.

General and specialized metabolites in peanut roots regulate arbuscular mycorrhizal symbiosis

Arbuscular mycorrhizae(AM)fungi form symbiotic associations with plant roots,providing nutritional benefits and promoting plant growth and defenses against various stresses.Metabolic changes in the roots during AM fungal colonization are key to understanding the development and maintenance of these symbioses.Here,we investigated metabolic changes in the roots of peanut(Arachis hypogaea L.)plants during the colonization and development of AM symbiosis,and compared them to uncolonized roots.The primary changes during the initial stage of AM colonization were in the contents and compositions of phenylpropanoid and flavonoid compounds.These compounds function in signaling pathways that regulate recognition,interactions,and pre-colonization between roots and AM fungi.Flavonoid compounds decreased by 25%when the symbiosis was fully established compared to the initial colonization stage.After AM symbiosis was established,general metabolism strongly shifted toward the formation of lipids,amino acids,carboxylic acids,and carbohydrates.Lipid compounds increased by 8.5%from the pre-symbiotic stage to well-established symbiosis.Lyso-phosphatidylcholines,which are signaling compounds,were only present in AM roots,and decreased in content after the symbiosis was established.In the initial stage of AM establishment,the content of salicylic acid increased two-fold,whereas jasmonic acid and abscisic acid decreased compared to uncolonized roots.The jasmonic acid content decreased in roots after the symbiosis was well established.AM symbiosis was associated with high levels of calcium,magnesium,and D-(+)-mannose,which stimulated seedling growth.Overall,specific metabolites that favor the establishment of AM symbiosis were common in the roots,primarily during early colonization,whereas general metabolism was strongly altered when AM symbiosis was well-established.In conclusion,specialized metabolites function as signaling compounds to establish AM symbiosis.These compounds are no longer produced after the symbiosis between the roots and AM becomes fully established.

Assessment of Photochemical Reflectance Index as a Tool for Evaluation of Chlorophyll Fluorescence Parameters in Cotton and Peanut Cultivars Under Water Stress Condition

The relationships between photochemical reflectance index （PRI） and the chlorophyll fluorescence parameters were examined to assess suitability of PRI as a remote-sensing tool for the chlorophyll fluorescence parameters. A greenhouse experiment was conducted using cotton and peanut crops under water stress condition. Five cotton and six peanut cultivars were grown using Andosole soil in pots maintained at two water levels; the control and water stress treatment （WS） of 100 and 50% of the daily transpiration, respectively. Higher non-photochemical quenching （NPQ） was exhibited by peanut than that of cotton by the water stress. On the other hand, the decreases of the actual quantum yield of photosystem II （△F/F＇m） and PRI by the water stress in cotton were larger than those in peanut. There were positively significant correlation coefficients between PRI and △F/F＇m in cotton at noon and in the afternoon including the control and WS. The correlations of PRI with NPQ were negatively significant at noon and in the afternoon for cotton, and in the afternoon for peanut. No clear relationship was found among these parameters in the morning probably due to the diurnal increase in global solar radiation. It was concluded that there would be a possibility to detect the effects of water stress on △F/F＇m and NPQ by PRI with some exceptions, although PRI could not note varietals differences in △F/F＇m and NPQ for each treatment.

Progress on genetic and genomic research for enhanced oil content and quality in peanut

Peanut or groundnut （ Arachis hypogaea L. ） is an important source of vegetable oil in the world. Genetic enhancement for high yield and high oil content has greatly contributed to enhanced pro-ductivity of peanut and increased supply of peanut oil. Further improving oil content and quality of peanut is still crucial for increasing productivity of arable land and market competitiveness of peanut oil. Based on investigation among the peanut germplasm accessions including wild Arachis species, the oil content could be as high as 65%. Heterosis has been observed for oil content in hybrids derived from diverse crossing parents. Segregates with enhanced oil content have been obtained by pyramiding different genes or alleles with major and minor additive effects. Improved testing techniques for oil content in breeding lines with reduced cost have accelerated breeding progress for high oil content. SSR markers associated with oil content have been identified by association and linkage analysis. The stability of oil content in peanut across seasons and locations is highly associated with cold tolerance, high nutrition efficiency and drought tolerance. Recent progress on improving fatty acids in particular marker assisted backcrossing breeding has contributed to higher quality peanut oil and other products. High oil peanut lines with im-proved resistance to aflatoxin production have been developed. The wild Arachis species would be of great value for peanut breeding in increasing oil content.

Infestation of Diatraea saccharalis(Fabr.)during Five Consecutive Cycles of Sugarcane Crop Succeeding Leguminous Crops

The area cultivated with sugarcane in Brazil is expanding, and increasingly using legumes cover crop in new deployed areas and in those which were reformed. Long-term trials were carried out in Piracicaba, Sao Paulo, Brazil, to evaluate the effect of leguminous green manure on the natural infestation of Diatraea saccharalis (Fabr.) during five cropping cycles of sugarcane “IAC 87-3396”. This study was carried out from October 1999 to October 2005 in Piracicaba, Sao Paulo, Brazil, at coordinates 22°42''S and 47°38''W, 560 m above sea level, in soil classified as Typic Paleudult. Green manure crops were assigned to five treatments: peanuts (Arachis hypogaea L.) (cultivar “IAC-Tatu” and “IAC-Caiapó”), sunn hemp (Crotalaria juncea L.) (cultivar “IAC 1”) and velvet-bean [Mucuna aterrima (Piper & Tracy) Holland] (cultivar “common”). A treatment without any green manure or weed was used as the control. The previous sugarcane crop was destroyed and the soil was prepared in a conventional way (using plow and harrow) before the leguminous crops were sowed. Thirty days after the sowing, weeds were removed and, 120 days later, the crops were cut and its biomass was deposited on the soil surface without incorporation. Experimental plots comprised five rows of sugarcane, each one measuring 10 m long and spaced 1.40 m apart. The sugarcane was harvested 18 months after biomass deposit, on October 25th, 2001 and its stalks were collected at intervals of approximately 12 months in: 07/09/2002, 08/01/2003, 11/07/2004 and 10/06/2005. At harvest, the masses were determined from a sample of sugarcane, cut from three rows (2 m long) in the center of the plot. The losses caused by the sugarcane borer were estimated by the intensity of the infestation by randomly observing and collecting sugarcane stems from each plot. A great reduction in the number of stalks and in the yield was noted proportionally to the intensity of the infestation of the borer in the sugarcane crop grown after the velvet-bean. The sugarcane-velvet-bean rotation should be regarded considering the intensity of sugarcane borer infestation. No influence of the other green manure crops was observed on the intensity of the infestations. However, the harvest seasons of the sugarcane seemed to influence the borer infestation.

Production of Peanut Intercropped with Forage Palm in Pernambuco State, Brazil

The greater use of crop fields by increasing plant population is an alternative to minimize losses to the farmers who use crop intercropped provides other benefits such as lower incidence of diseases and pests, and better exploitation and enrichment agroecosystem. The objective of this study was to evaluate the effect of the population of peanuts with and without inoculation and intercropping with forage palm on growth, productivity and biomass peanut. The treatments consisted of different populations of peanut (one, two and three rows of peanut row spacing of two meters of cactus pear), with and without inoculant, analyzed in subdivided plot scheme (3 × 2), totalizing six treatments in a randomized block design with four replications. The variables evaluated were plant height, number of branches, shoot dry mass and root nodule number, nodule dry weight, biomass productivity and legumes. The largest population of plants in number of lines of peanut intercropped with forage cactus promotes greater total biomass production and yield of beans and does not alter the height, branch number, dry mass of aerial part roots and nodules. The inoculant application did not result in to increased productivity biomass and growth of peanut plants, their use is not indicated. The planting of a row of peanuts between the rows of palm is not economically recommended.

Molecular characterization of a peanut variety and its derivatives based on SSR and COP analysis

Despite the economic importance of the peanut, no studies have been carried out to deter-mine the correlation between genetic distances based on molecular markers and on coefficient of parentage （COP） data. In this study, simple sequence repeat （SSR） and pedigree data were used to assess the ge-netic distance between the Fuhuasheng variety and its derivative cultivars. A total of 39 SSR polymor-phism primers were used, and 151 bands were obtained, with an average of 2. 04 bands in each primer. The genetic SSR - based distance （GD） values ranged from 0. 02 to 0. 81, while the COP - based GD ranged from 0. 25 to 0. 98. Certain Fuhuasheng loci displayed higher transmission rates. These loci or nearby chromosomal regions might be associated with desirable traits in Fuhuasheng variety, thus being frequently selected in breeding programs. Therefore, it can be suggested that COP analysis should be the preferred method for estimating genetic diversity invarieties with available complete pedigree information and parents. In this case, marker analysis would provide the best estimations.

不同播种方式下高油酸花生生长性状及产量的表现

以高油酸花生(Arachis hypogaea L.)品种中花215、中花28和中花34为试验材料,采用小区试验对比的方法研究不同播种方式(单粒播种、双粒播种、单双粒间播)对高油酸花生生长性状及产量的影响。结果表明,不同播种方式对高油酸花生生长性状及产量均产生一定影响,单粒播种更有利于花生单株生长发育,相比其他两种播种方式,能显著提高叶片SPAD值、单株结果数、单株生产力和干果产量;单双粒间播和双粒播种的干果产量无显著差异;3个高油酸品种单粒播种的经济效益最高,单双粒间播次之,双粒播种最低;相同播种方式下,中花215的产量和经济效益最高,中花34次之,中花28最低。

花生巢式群体的脂肪含量遗传分析

巢式群体可以利用多个亲本解析复杂性状的遗传机制。本研究利用1个共同亲本与6个基础亲本所配置巢式组合F_(2:3)家系的种子脂肪含量数据,分析了花生脂肪含量的遗传模型,旨在探明不同的基础亲本组合中脂肪含量性状的遗传差异,为制定脂肪含量遗传改良的亲本选配和后代选择策略提供依据。6个组合的共同亲本为高脂肪含量的普通型大果品种豫花15号,其他6个基础亲本为不同脂肪含量和不同植物学类型的品种。结果表明,在不同杂交组合中脂肪含量的遗传模式有所不同,6个组合分别符合无主基因模型、1对主基因加性显性模型和2对主基因等显性模型3种遗传模式。各种遗传效应的估计值也各不相同,主基因遗传力从32%到80%,说明不同杂交组合中,控制脂肪含量的基因位点差异及其重组和分离方式不同。高脂肪含量双亲杂交后代的高脂肪含量个体较多,但主基因遗传力较低,不宜在早代实施表型选择;双亲脂肪含量差异较大的后代脂肪含量变异幅度更大,能够选择到不同脂肪含量的类型。本研究也表明,巢式组合具有较丰富的脂肪含量变异类型,揭示出脂肪含量性状遗传的复杂性和多基因调控的特点,为较全面地了解脂肪含量的遗传提供了基础。该巢式群体也将有助于进一步开展脂肪含量的QTL定位研究。

花生抗网斑病品种筛选及抗病性与产量损失的关系

花生茎点霉(Phoma arachidicola Marasas Pauer&Boerema)引起的花生网斑病在生产上普遍流行、危害严重,2018-2019年通过2年的田间接种试验对国内65份花生种质资源网斑病抗性进行测评。以采自山东莱西的病原菌菌株Wb2制备105/mL的孢子悬浮液喷洒于花生叶片表面进行接种,对照区喷施50%咯菌腈WP防治花生网斑病菌。结果表明,65份种质中,抗病(resistant,R)资源8份,占鉴定资源总数的12.3%;中度抗病(moderately resistant,MR)资源9份,占比13.8%;感病(susceptible,S)资源37份,占比56.9%;高度感病(high susceptible,HS)资源11份,占比16.9%。测定抗病性不同资源产量损失差异,结果表明,花生网斑病菌对花生产量影响显著,产量损失率随抗病性的降低而升高。本研究为花生抗网斑病育种提供抗源材料,并为病害产量损失评估提供理论依据。

Advances in Photosynthetic Characteristics of Peanut

Peanut is an important oil crop and economic crop in China. In this pa- per, the status and advances of the factors affecting peanut photosynthesis, such as light intensity, temperature, carbon dioxide concentration, water deficit and fertility situation, the relationship between peanut photosynthetic rate and different cultivation system, and the relationship between peanut photosynthetic rate and varieties yield potential, were deeply reviewed, expecting to provide references for peanut variety breeding with high photosynthetic efficiency and high yield potential.

巢式杂交群体的花生荚果性状遗传模型分析

采用主基因+多基因遗传模型,对巢式群体的5个组合的F2家系的荚果性状进行了遗传模式解析,以期了解巢式杂交群体的荚果性状遗传变异特点。结果表明,巢式杂交群体具有丰富的荚果性状的变异类型,荚果长、宽和百果重在5个组合中的最小值至最大值变异幅度分别为(14.30~22.09)mm~(38.36~45.12)mm、(7.06~10.47)mm~(17.13~22.74)mm和(62.41~94.38)g~(266.75~364.00)g。荚果长与荚果宽、荚果表面积、荚果表面周长、百果重的相关性均极显著,与荚果长宽比的相关性较小;荚果宽与荚果表面积、荚果表面周长、百果重存在正相关,与荚果长宽比存在负相关。不同杂交组合的不同果型性状的遗传模式均有差异,最佳遗传模型为两对主基因加性-显性模型和两对主基因加性-显性-上位性模型;主基因遗传力22.79%~91.62%,不同群体中的基因效应值各不相同,表明多等位基因或非等位基因的不同遗传效应以及遗传背景差异对荚果性状的影响。本研究为利用NAM群体开展荚果性状QTL定位及分子标记开发、为专用型花生新品种选育提供了材料基础和理论依据。

自然富硒花生种质资源的筛选

在天然富硒旱地上,通过田间试验对20个花生(Arachis hypogaea L.)品种的子粒进行硒含量检测。结果表明,不同花生品种子粒的硒含量差异极显著,变幅为0.041~0.163 mg/kg,不同花生品种对土壤硒的利用效率存在极显著差异,变幅为6.7%~26.8%,共有15个品种达到了富硒水平,富硒率高达75%,其中天府23、大城花生、天府18、天府9的硒含量较高,硒利用效率也均大于20%,表现出较强的富硒能力,可作为高富硒花生种质资源用于后期富硒花生新品种的选育。试验表明不同花生品种的富硒能力差异较大,进行花生富硒品种筛选对开发利用宜春市天然富硒土壤资源和富硒花生产业具有重要意义。

莱阳市花生产业概况、问题及发展建议

花生(Arachis hypogaea L.)是重要的油料作物和经济作物,具有产量高、经济效益好的特点,在山东省莱阳市广泛种植。分析莱阳市花生的种植优势、企业优势、品质优势、项目优势,厘清该产业发展过程中存在的问题,提出针对性的对策与建议,为推动莱阳市花生产业高质量发展提供参考。