Molecular fingerprinting of hybrids and assessment of genetic purity of hybrid seeds in rice using microsatellite markers

Microsatellite markers were used for fingerprinting of hybrids, assessing variation within parental lines and testing the genetic purity of hybrid seed lot in rice. Ten sequence tagged microsatellite sites (STMS) markers were employed for fingerprinting 11 rice hybrids and their parental lines. Nine STMS markers were found polymorphic across the hybrids and produced unique fingerprint for the 11 hybrids. A set of four markers (RM 206, RM 216, RM 258 and RM 263) differentiated all the hybrids from each other, which can be used as referral markers for unambiguous identification and protection of these hybrids. Cluster analysis based on Jaccard's similarity coefficient using UPGMA grouped the hybrids into three clusters. Within the cluster all the hybrids shared a common cytoplasmic male sterile line as female parent. The genetic similarity between the hybrids ranged from 0.33 to 0.92 with an average similarity index of 0.63. The analysis of plant-to-plant variation within the parental lines of the hybrid Pusa RH 10, using informative markers indicated residual heterozygosity at two marker loci. This highlights the importance of STMS markers in maintaining the genetic purity of the parental lines. The unique value of the restorer gene linked marker for testing the genetic purity of hybrid seeds is demonstrated for the first time.     

Prediction of heterosis for grain yield in rice using 'key' informative EST-SSR markers

This study was undertaken to assess the comparative potential of 25 Expressed Sequence Tag derived simple sequence repeats (EST-SSRs) and 25 genomic SSRs in the prediction of grain yield heterosis using a set of nine cytoplasmic male sterile (CMS) lines and 32 restorer lines of rice. EST-SSRs and genomic SSRs exhibited an average Polymorphism Information Content value of 0.37 and 0.45, respectively. The coefficient of marker polymorphism among parental lines with respect to a set of hypervariable EST and genomic SSRs was correlated with standard heterosis for grain yield of six public bred rice hybrids. EST-SSRs gave a better correlation (r = 0.75) as compared with genomic SSRs (r = 0.09). When 10 'key' informative EST-SSR markers which showed a higher positive correlation with grain yield heterosis were validated in a new set of 14 experimental hybrids, the markers exhibited a higher correlation (r = 0.79), indicating the predictive value of these EST-SSRs. We recommend these 10 'key' informative EST-SSR markers for analysis of genetic diversity of parental lines and prediction of heterosis in hybrid rice breeding programmes.     

Relationship between molecular marker heterozygosity and hybrid performance in intra- and interspecific hybrids of cotton

The present study was conducted to investigate the relationship between parental molecular marker diversity and hybrid performance in both intra‐ and interspecific hybrids of cotton to evaluate the feasibility of predicting hybrid performance using molecular markers. Three cytoplasmic male sterile (CMS) lines were crossed with 10 restorer lines to produce 22 F₁ hybrids during 2003. Of 22 F₁s, 14 hybrids were intraspecific (Gossypium hirsutum × G. hirsutum) and eight interspecific (G. hirsutum × G. barbadense). These 22 F₁ hybrids and their parents were evaluated for yield and fibre quality traits at Zhejiang University, Hangzhou, China during 2004 and 2005. Genetic distances (GD) among the parents were calculated from 56 random‐amplified polymorphic DNAs (RAPD) and 66 simple sequence repeat (SSR) marker data, and their correlation with hybrid performance and heterosis were analysed. The parents could be discriminated into G. hirsutum and G. barbadense clusters by cluster analysis based on both RAPD and SSR markers data. The correlation (r = 0.503, P ≤ 0.05) was calculated between GD_rapd (GD based on RAPD markers) and GD_ssr (GD based on SSR markers). Correlation of GD with hybrid performance and heterosis differed considerably between intra‐ and interspecific hybrids. The correlation between GD and hybrid performance was non‐significant for most of traits within the hybrids of G. hirsutum species. However, it was significantly and positively correlated for fibre length, fibre strength and elongation in interspecific hybrids. The relationship between GD and heterosis was observed to be positively significant for boll weight within hybrids of G. hirsutum with significant and negative correlations for fibre length and elongation. In conclusion, the power of predicting hybrid performance using molecular markers in cotton is low. But, the relationship between SSR marker heterozygosity and hybrid performance can be used to predict fibre length during interspecific hybrid cotton breeding.     

Development of a DNA marker for distinguishing CMS lines from fertile lines in rice (Oryza sativa L.)

The main objective of the present study was to identify mitochondrial DNA based marker, which can distinguish male sterile and fertile counterparts of the cytoplasmic male sterile (CMS) lines used in production of rice hybrids. Amplified fragment length polymorphism (AFLP) analysis in CMS lines: IR58025A & IR62829A and their respective maintainers: IR58025B & IR62829B identified a polymorphic DNA fragment of about 510 bp size that was present in both CMS (A) and absent in their maintainer (B) lines. Sequencing followed by database analysis of the polymorphic fragment indicated about 97% similarity with mitochondrial NADH gene subunits of rice, maize and wheat. Based on the variable sequence regions, a site specific primer pair (BF-STS-401) was designed. PCR analysis showed that BF-STS-401 could amplify a strong band of 464 bp size in CMS and a faint band of the same size in maintainer line. To act as a positive control and avoid possible errors in PCR, BF-STS-401 was multiplexed with a new primer pair (BF-STS-402), derived from mitochondrial atp9 subunit of rice, producing monomorphic amplification indiscriminately in both CMS and maintainer lines. Both the primer pairs in combination clearly differentiated CMS lines from their corresponding maintainer lines. This primer combination was validated in a set of diverse genotypes consisting of different sources of CMS lines, restorer lines, hybrids, varieties and mixed samples from private seed companies. Our results suggested that the multiplex primer pairs developed in this study can be effectively utilized to assess the genetic purity in commercial seed lots of CMS lines and hybrids of rice.     

杂交粳稻亲本产量性状优异配合力的标记基因型筛选

提高杂交粳稻竞争优势的关键是改良其恢复系产量性状的配合力。为使之更富成效,选用115个SSR引物扩增6个粳稻BT型不育系和12个恢复系的标记基因型,并按NCII遗传设计配制72个F1组合,分析18个亲本的单株日产量、单株有效穗数、每穗总粒数、每穗实粒数、千粒重5个性状的配合力,结合亲本SSR分子标记数据和性状配合力数据筛选了5个性状优异配合力的标记基因型。结果发现20个SSR标记基因型与亲本产量及其构成性状配合力显著相关。其中8个与亲本单个性状配合力相关;6个同时与亲本2个性状配合力相关;4个同时与亲本3个性状配合力相关;2个同时与亲本4个性状配合力相关。同时与多个性状配合力相关的标记基因型,对各性状的作用方向有正有负。RM152-165/170是单株日产量和单株有效穗数优异配合力效应最大的标记基因型,可使F1的相应性状值增加20.6%和12.7%。优异配合力的标记基因型可直接用于粳稻恢复系配合力的分子标记辅助改良。     

A simple method to control the seed purity of japonica hybrid rice varieties using PCR-based markers

Cytoplasmic male sterility (CMS) by the cms‐bo cytoplasm and its restoration by the nuclear restorer gene, Rf‐1, are used for seed production of japonica hybrid rice varieties. To produce pure hybrid seeds, a prerequisite is to properly manage the seed purity of parental lines, especially CMS lines. In this study, three dominant polymerase chain reaction (PCR)‐based markers (M1, M2 and M3) were developed to detect mutual contamination in seed batches of CMS lines, maintainer lines, restorer lines and hybrids. M1 detected the mitochondrial sequence that was present in the cytoplasm of common japonica varieties and absent in the cms‐bo cytoplasm. M2 and M3 detected the chromosomal sequence related to the Rf‐1 allele in restorer lines and the rf‐1 allele in common japonica varieties, respectively. By the strategic use of these markers, japonica hybrids and their parental lines could be efficiently distinguished from each other. Furthermore, sensitivity tests for the three markers with a series of crude DNA samples prepared from polished grains demonstrated that these markers could detect one contaminating grain among 500 or 1000 grains. Therefore, the bulk PCR analyses with the markers developed here probably make it possible to control the seed purity of japonica hybrids properly by selecting appropriate seed batches of their parental lines quickly and efficiently.     

江淮稻区杂交粳稻骨干亲本产量性状配合力的SSR标记位点鉴定

鉴定杂交粳稻亲本产量性状配合力的标记位点有助于利用分子标记辅助选择技术改良亲本配合力、提高杂交粳稻竞争优势水平。利用9个粳稻BT型不育系和10个恢复系, 按照北卡罗林那设计II (North Carolina Design II)配制90个F₁组合,在南京和盱眙两个环境下种植,测定了各亲本产量性状的配合力和SSR分子标记基因型;结合二者数据鉴定了6个产量性状配合力的标记位点。结果表明,在两个环境下综合评价配合力较优的不育系是BT-18A和武羌A,恢复系是C418。与亲本单株有效穗数、每穗总粒数、每穗实粒数、结实率、千粒重和单株日产量性状配合力显著相关的SSR标记位点,南京环境下分别检测到8、13、11、6、6和2个;盱眙环境下分别检测到12、21、8、15、10和7个;2个环境下都检测到的分别有4、11、5、3、5和1个。标记位点杂合基因型显示正向优势的,南京环境下占74% (34/46);盱眙环境下占53% (39/73)。2个环境下都检测到的标记位点中,有3个各自与3个产量性状配合力共相关;另有3个各自与2个产量性状配合力共相关;其余的只与单个产量性状配合力显著相关。数据库检索发现两环境下都检测到的标记位点中,有10个其附近存在控制相应性状的基因/QTL。讨论了利用鉴定出的标记位点改良粳稻恢复系产量性状配合力的策略。     

Assessment of purity of rice CMS lines using cpDNA marker

AFLP technique was used to analyse the polymorphism between rice cytoplasmic male sterility (CMS) line Jin2A and its maintainer Jin2B. A stable differential band was discovered, and sequence analysis showed that Jin2A contained a more tandem repeat of 6 base pairs (AGAAAA) than Jin2B. Further studies confirmed that the diversity came from cpDNA and occurred at three kinds of abortive cytoplasmic genotypes. Accordingly, specific primers were designed and utilized to assess the purity of rice CMS lines during multiplication with pollen fertility and seed setting rates of bagged panicles as control. The result indicated that this cpDNA locus could be utilized to precisely distinguish maintainer plants from rice CMS lines. PCR analysis was consistent with that from Grow-out test in CMS line seed purity assessment during multiplication, despite it was helpless in distinguishing F₁ hybrids from CMS lines due to similar cytoplasms. Because of fewer hybrid and more maintainer off-plants, this cpDNA locus was still appropriate for seed purity assessment of rice CMS line during multiplication. This is first report that a marker on cpDNA could be utilized to assess the genetic purity of rice CMS lines with three abortive cytoplasmic genotypes.     

Molecular mapping of a fertility restorer gene for cytoplasmic male sterility in soybean

In this study, we report the mapping of the Rf locus in soybean by microsatellite simple sequence repeat (SSR) genetic markers. A cross was made between cytoplasmic male sterility (CMS) line JLCMS82A and restorer line JIHUI 1 based on the DNA polymorphisms revealed by 109 SSR markers. A F₂ population derived from a single F₁ plant containing 103 individuals was used for mapping the Rf locus. The Rf gene of JIHUI 1 gametophytically restores male fertility to JLCMS82A. Fertile and semi-fertile DNA bulks and parental DNAs were screened with 219 SSR markers, and Satt215 which was previously mapped to soybean LG J, was found linked to the Rf gene. Five additional polymorphic SSR markers from LG J were used for analysis and a regional linkage map around the Rf locus was established. SSR markers, Sctt011 and Satt547, flanked the Rf locus at 3.6 cM and 5.4 cM, respectively. The availability of these SSR markers will facilitate the selection of restorer lines in hybrid soybean breeding.     

Mapping of male sterility gene ms10 in chilli pepper (Capsicum annuum L.)

Most of the hybrid seed in chilli are produced manually, but the use of male sterility (MS) can reduce the cost of hybrid seed production. MS‐12, a nuclear male‐sterile (NMS) line developed at Punjab Agricultural University, Ludhiana (India), has been utilized to develop commercial F₁ hybrids. A recessive gene, designated as ms10, governs MS in MS‐12. Due to recessive gene control, development of new NMS lines incorporating ms10 gene is tedious and time‐consuming. We identified SSR markers AVRDC‐PP12 and AVRDC_MD997* linked to the ms10 gene. A total of 558 primer pairs were screened following bulked segregant analysis (BSA). Linkage analysis in 210 F₂ plants indicated that the two SSR markers were linked to the ms10 gene and the marker AVRDC‐PP12 was closest to the gene at 7.2 cM distance. The marker was mapped to chromosome 1 at genome position 175 694 513 to 175 694 644. Until more closely linked markers are developed, the marker AVRDC‐PP12 would facilitate transfer of ms10 gene through marker‐assisted selection (MAS). Fine mapping would lead to cloning of the ms10 gene.     

利用SSR技术快速准确鉴定杂交玉米种子纯度

利用一种快速、廉价的DNA提取方法,提取了两个玉米杂交种及其亲本的DNA用于SSR分子标记分析。分析结果显示,10对被分析的引物中有4对在两个杂交种双亲之间显示出多态性,可以用于相应的杂交种纯度分析。另外,分别利用同工酶技术和SSR分子标记技术同时分析了来自这两个杂交种的种子样品的纯度,分析发现,对于一个杂交种,两种方法都能可以鉴定,并且两者检测的结果是一致的,但是对于另外一个杂交种,由于其父母本非常接近,同工酶不能将其区分。因此,这些结果显示SSR分子标记技术可以很好地用于杂交玉米种子的纯度鉴定,即使是这个杂交种是来自两个非常接近的自交系的。     

Tagging of four fertility restorer loci for wild abortive—cytoplasmic male sterility system in rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.) using microsatellite markers

Tagging of restorer genes for wild abortive (WA) CMS source by studying a 222 individual plants from a F₂ population of a cross between IR58025A × IR42686R. The restorer line IR42686R that was used in this study had been previously derived through random mating composite population (RMCP) involving 12 parents facilitated by IR36 genetic male sterility. Four Rf genes were tagged to simple sequence repeats (SSR) markers on chromosomes 1, 7, 10, 12 by recessive class analysis. The recombination frequency between a positive marker and Rf locus was calculated using maximum likelihood estimator assuming that all the 46 extremely sterile individual plants were homozygous at the targeted Rf locus. The recombination frequency between the marker and the restorer trait were converted to genetic distances using Kosambi function. A new Rf locus designated as Rf7 on chromosome 12 was found to be linked to RM7003 at a genetic distance of 13.3 cM (LOD 6.12). We report here first, a new molecular marker (RM 6344) linked to Rf4 locus on chromosome 7 that was previously mapped by trisomic analysis. RM443 and RM315 were flanking the Rf3 gene at a genetic distance of 4.4 (LOD 10.29) and 20.7 cM (LOD 3.98) on chromosome 1, respectively. The Rf6 was flanked on both side with SSR markers RM258 and RM591 at a genetic distance of 4.4 (LOD 10.29) and 23.3 cM (LOD 3.39) located on chromosome 10. The random mating composite population is an excellent breeding approach to develop superior restorer lines and for pyramiding different Rf genes of different CMS systems. Rf genes tagged with closely linked SSR markers would be facilitating marker assisted selection (MAS) in hybrid rice breeding program by reducing time and workload for identifying potential restorers. L. Bazrkar and A. J. Ali equally contributed to this work.     

利用SSR分子标记划分杂交籼稻亲本群的研究

杂种优势群和杂种优势利用模式可以为植物的杂种优势利用提供重要信息。与玉米相比,水稻杂种优势群的研究相对薄弱。本研究利用72对SSR引物对我国47个杂交籼稻骨干亲本进行了类群划分。共检测出328个等位基因变异,每对引物检测等位基因2~13个,平均4.56个;引物的多态性信息量（PIC）范围为0.120~0.878,平均值为0.567。材料之间的遗传相似系数(GS)范围在0.633~0.928,平均为0.741。用类平均法（un-weighted pair-group method using an arithmetic average）将亲本材料划分为保持系群、温敏核雄性不育系群、恢复系群,其中保持系群和恢复系群又各分为3个亚群,分群结果基本符合系谱信息。并结合杂交水稻生产实践提出了7种杂种优势组合配组模式。分子标记是一种划分杂交籼稻亲本群的有效途径,本研究结果对杂交籼稻的亲本选配具有一定指导意义。     

Influence of Male Sterile Cytoplasms on the Physico-Chemical Grain Quality Traits in Hybrid Rice (Oryza Sativa L.)

Summary Possible unfavourable influence of sterility inducing cytoplasm on physico-chemical grain quality traits in rice hybrids is one of the important concerns hindering the large-scale adoption of hybrid rice technology. Produce from 23 pairs of CMS line × restorer (AF₁) and maintainer line × restorer (BF₁) cross combinations carrying different cyto-sterile sources (WA, ARC, Mutagenized IR 62829B and Kalinga I) was compared for various grain and cooking quality traits. The milling recovery in rice hybrids was not influenced by the sterile cytoplasm. For kernel dimensions before and after cooking there were both favourable and unfavourable cytoplasmic effects, which varied in magnitude depending upon the sterile cytoplasm and parental combinations. Similar results were obtained for kernel elongation and gelatinization temperature. The most widely used WA cytoplasm had minimum instances of unfavourable influence. In general, the cytoplasmic influence was found to be highly cross-specific and depended on the nuclear background of CMS line and fertility restorer. Availability of alloplasmic CMS lines carrying different cyto-sterile sources in the same nuclear background would help in ascertaining the cytoplasmic influence in a more comprehensive manner.     

杂交粳稻亲本米质性状优异配合力的标记基因型鉴定

杂交粳稻米质整体水平不如常规粳稻也是限制杂交粳稻广泛种植的原因之一。本研究选用115个SSR引物扩增6个粳稻BT型不育系和12个恢复系的标记基因型,并分析72个F₁组合谷粒长、谷粒宽、糙米率、精米率、整精米率、垩白米率、垩白度、糊化温度、胶稠度和直链淀粉含量10个米质性状的配合力,结合亲本SSR分子标记数据和性状配合力数据筛选了10个米质性状优异配合力的标记基因型。结果共鉴定出30个SSR标记基因型与亲本10个米质性状配合力显著相关,其中25个与亲本米质性状不良配合力相关,5个与优异配合力相关。标记基因型RM263-175/180和RM444-230/240可以使F₁整精米率分别提高3.2%和2.5%。RM3-120/150可以使F₁谷粒长缩短2.4%,RM444-180/240可以使F₁谷粒宽增加2.1%。RM428-273/294可以使F₁植株上的杂交稻米直链淀粉含量减少7.0%。有8个标记基因型同时也影响产量性状配合力。RM3-120/150同时可以使F₁的每穗总粒数和每穗实粒数分别增加15.9%和10.9%。RM1211-150/160可使F₁的糙米率和精米率分别减少0.9%和1.1%,同时使F₁的每穗总粒数和每穗实粒数分别增加21.8.%和20.4%。RM23-150/160可使F₁的垩白米粒率和垩白度分别增加44.1%和45.7%,同时使F₁的单株日产量和每穗总粒数分别增加11.2%和11.6%。这些结果可用于指导亲本米质性状和产量性状配合力的分子标记辅助改良以及未来杂交粳稻组合配置中的亲本选配。     

Relationship between molecular marker heterozygosity and hybrid performance in intra- and inter-subspecific crosses of rice

The objective of this study was to investigate the relationship between molecular marker diversity and heterosis in both intra-and inter-sub-specific hybrids of rice to evaluate the feasibility of predicting hybrid performance using molecular markers. Eleven elite lines were intermated resulting in a diallel set including 10 indica × indica, 15 japonica × japonica and 30 indica × japonica crosses. The F₁ hybrids and parents were evaluated for agronomic performance in a replicated field trial. The parental lines were tested for DNA polymorphisms with 113 restriction fragment length polymorphism (RFLP) probes covering the 12 rice chromosomes. Inter-subspecific crosses showed better performance and higher heterosis than intrasubspecific hybrids. Correlations of marker heterozygosity with hybrid performance and heterosis differed considerably between the two subspecies; they were higher in crosses within japonica subspecies than within indica subspecies. Very little correlation was detected in intersubspecific crosses. It was concluded that relationships between marker heterozygosity and hybrid performance were complex owing to germplasm diversity and the complexity of the genetic basis of heterosis. The implications of the results in predicting hybrid performance using molecular markers are discussed.     

Relationship of genetic distance and hybrid performance in hybrids derived from a new photoperiod-thermo sensitive male sterile wheat line 337S

Heterosis has been exploited in many crops and made a significant contribution to the world food supply. Genetic distance (GD) is one of valuable criteria for selecting parents in hybrid breeding. The objectives of this study were to estimate GD between a novel photoperiod-thermo sensitive male sterile wheat line 337S and several common cultivars using simple sequence repeat (SSR) markers, and to evaluate the relationship of GD and heterosis. The line 337S, as a maternal parent, was crossed with 16 common wheat varieties to produce 16 hybrids. Eight agronomic traits were investigated for the parents and F1 hybrids. Mid-parent heterosis (MPH) and specific combining ability (SCA) were calculated. GD and cluster analysis were performed among the 17 parents. The correlations of GD with F1 performance, MPH and SCA were analyzed. The results showed that GD estimates among the 17 parents averaged 0.42 with a range from 0.26 to 0.57, and the averaged GD between 337S and the 16 parental cultivars was 0.44. Cluster analysis separated the 17 parents into three main groups. SSR markers were useful in analyzing genetic divergences among the 17 parental lines. However, GD based on SSR markers poorly correlated with F1 performance, MPH and SCA. Thus, GD revealed by SSR markers can not provide a reliable prediction in practical wheat hybrid breeding.     

Prediction of hybrid performance based on the genetic distance of parental lines in two-line rice (Oryza sativa L.) hybrids

A total of six TGMS (thermosensitive genic male sterile lines) and nine pollinator lines were subjected to molecular characterization using 48 genome-wide SSR (simple sequence repeat) markers. Cluster analysis revealed clear differentiation among the TGMS lines according to their source of origin. The SSR-based genetic distance between the hybrids of the parental lines ranged from 0.36 to 0.79 suggesting a high degree of genetic divergence. Among a set of 54 hybrids generated using parental lines, 32 showed better parent heterosis (+21.7%) while 19 showed mid-parent heterosis (+15.0%). For the trait yield per plant genetic distance (GD) was negatively correlated with F₁ performance (r = ?0.202), mid-parent heterosis (r = ?0.325*; P < 0.05), and better parent heterosis (r = ?0.261), while it was positively correlated with specific combining ability (r = 0.042). Based on the grouped genetic distance (GGD), the hybrid combinations were divided into four groups. The GGD showed linear correlation with hybrid performance within the group (GGD = 40–50: r = ?0.07; GGD = 70–80: r = 0.32). This information can be utilized in the development of higher yielding, two-line rice hybrids through selection of intermediately diverse parental lines using GGD.     

水稻广亲和广谱型恢复系SWR78的选育

利用野败（WA）型、红莲（HL）型、包台（BT型）3种细胞质的同核异质粳稻广亲和不育系苏秋A与19个籼、粳恢复系测交,筛选出对以上3种细胞质不育系均具正常恢复力的广谱型恢复系6078。通过回交,将广亲和品种Lemont的广亲和基因导入6078,育成广亲和广谱型恢复系SWR78。经测交鉴定,SWR78对WA型、印水（ID）型、HL型、BT型4种细胞质的10个籼、粳不育系均具正常恢复力。     

Combining ability,heterosis and genetic diversity in tropical maize (<Emphasis Type="Italic">Zea mays</Emphasis> L.) under stress and non-stress conditions

Drought and low soil fertility are considered the most important abiotic stresses limiting maize production in sub-Saharan Africa. Knowledge of the combining ability and diversity of inbred lines with tolerance to the two stresses and for those used as testers would be beneficial in setting breeding strategies for stress and nonstress environments. We used 15 tropical maize inbred lines to (i) evaluate the combining ability for grain yield (GY), (ii) assess the genetic diversity of this set of inbred lines using RFLP, SSR, and AFLP markers, (iii) estimate heterosis and assess the relationship between F₁ hybrid performance, genetic diversity and heterosis, and (iv) assess genotype × environment interaction of inbred lines and their hybrids. The F₁ diallel hybrids and parental inbreds were evaluated under drought stress, low N stress, and well-watered conditions at six locations in three countries. General combining ability (GCA) effects were highly significant (P < 0.01) for GY across stresses and well-watered environments. Inbred lines CML258, CML339, CML341, and CML343 had the best GCA effects for GY across environments. Additive genetic effects were more important for GY under drought stress and well-watered conditions but not under low N stress, suggesting different gene action in control of GY. Clustering based on genetic distance (GD) calculated using combined marker data grouped lines according to pedigree. Positive correlation was found between midparent heterosis (MPH) and specific combining ability (SCA), GD and GY. Hybrid breeding program targeting stress environments would benefit from the accumulation of favorable alleles for drought tolerance in both parental lines.