Introgression of <Emphasis Type="Italic">Gossypium barbadense</Emphasis> L. into Upland cotton germplasm RMBUP-C4S1

Gossypium barbadense L. cotton has significantly better fiber quality than Upland cotton (G. hirsutum L.); however, yield and environmental adaptation of G. barbadense is not as wide as Upland. Most cotton in the world is planted to Upland cultivars. Many attempts have been made, over a considerable number of years, to introgress fiber quality alleles from G. barbadense into Upland. However, introgression barriers, primarily in the form of interspecific incompatibility, have limited these traditional approaches. The use of chromosome substitution lines (CSL) as a bridge should provide a more efficient way to introgress alleles from G. barbadense into Upland. We crossed 18 G. barbadense CSL to three cultivars and developed a random mated population. After five cycles of random mating followed by one generation of self-pollination to increase the seed supply, we grew the random mated population and used 139 G. barbadense chromosome specific SSR markers to assess a random sample of 96 plants for introgression. We recovered 121 of 139 marker loci among the 96 plants. The distribution of the G. barbadense alleles ranged from 10 to 28 alleles in each plant. Among the 96 plants we found individual plants with marker loci from 6 to 14 chromosomes or chromosome arms. Identity by descent showed little relatedness among plants and no population structure was indicated by a heat map. Using CSL we were able to develop a mostly Upland random mated population with considerable introgression of G. barbadense alleles which should be useful for breeding.     

Hybridization between cotton and Malvaceae species as a tool for production of partial interspecific aneuploid cotton plants

Cotton although is an autogamous species could be cross pollinated under favorable climate conditions and/or in the presence of pollinators. The coexistence of cotton with Malvaceae species raises questions on the possibility pollen to be exchanged among Malvaceae species and on the resulted consequences. The present work was undertaken to evaluate the in situ response of cotton flowers (G.hirsutum L, G. barbadense L.) and their F₁ interspecific hybrids when are artificially pollinated with Malva sylvestris L., Hibiscus syriacus L. and Abelmoschus esculentus Moench. Furthermore, an in vitro protocol was attempted to support embryos’ growth in order to produce viable progenies originating from crosses between cotton and the aforementioned Malvaceae species. The obtained results gave evidence that pollen from the above Malvaceae species stimulated cotton ovaries without successful hybridization. The interaction between pollen and cotton’s stigmas was higher at early stages when M. sylvestris was used as pollinator but in crosses with H. syriacus and A. esculentus more ovules were activated as revealed by the percentage of carpodesis and life-time of bolls onto maternal plants. Only crosses between cotton with okra produced cotton seeds under in situ conditions. The in vitro embryo-ovule culture protocol, used, increased the number of regenerated cotton plants, especially in crosses among F₁ interspecific cotton hybrids and A. esculentus. In this case, regenerated plants were recombinant aneuploids, combining traits from both cotton species. This novel cotton germplasm possessing unique chromosome rearrangements, at aneuploid level could be proved useful after cytogenetic, molecular or QTL genetic analysis referring to important agronomic traits.     

Genetic relationships among resynthesized,semi-resynthesized and natural <Emphasis Type="Italic">Brassica napus</Emphasis> L. genotypes

The allopolyploidization event that created cultivated oilseed rape Brassica napus L, followed by intense breeding, reduced its genetic diversity. Resynthesized (RS) B. napus L. obtained by interspecific hybridization between genotypes of B. rapa L. and B. oleracea L. can be a valuable source for broadening genetic diversity in cultivated oilseed rape. In this study, we determined the extent of DNA polymorphism among natural accessions of oilseed rape, resynthesized B. napus, their parental species and double-low quality semi-RS lines carrying the Rfo gene. Using 10 selected primer combinations, 522 polymorphic AFLP markers were scored in the complete set of 100 Brassica sp. To detect relationships between these genotypes, a cluster analysis was performed using the Jaccard’s distance. Resynthesized allopolyploids clustered directly between their diploid parents. Cultivated accessions of oilseed rape created a compact group away from resynthesized allopolyploids as well as semi-RS lines. The natural oilseed rape group, which consists of 49 cultivars and breeding lines of oilseed rape, is characterized by lower genetic diversity than the group of 33 accessions of resynthesized oilseed rape, and the analysis showed that the double-low quality semi-RS lines represent a specific genetic variation of B. napus. The de novo resynthesized B. napus lines and the semi-RS lines of double-low quality generated from them, provide a significant opportunity for enrichment the gene pool of oilseed rape.     

Morph-physiological responses of cotton interspecific chromosome substitution lines to low temperature and drought stresses

Limited knowledge about genetic and physiological traits associated with drought and low temperature stresses and narrow genetic diversity in Upland cotton (Gossypium hirsutum L.) are serious impediments in its genetic improvement. The objectives of this research were to determine the genetic and physiological traits associated with drought and low temperature effects and to identify chromosomal effects on these traits using chromosome substitution (CS) lines from three alien species of Gossypium, G. barbadense, G. tomentosum, and G. mustelinum, respectively. Two experiments were conducted to study low temperature and drought stress effects during seedling emergence and early growth stages in 21 cotton CS-lines with parent, Texas Marker (TM)-1. In Experiment I, plants were grown at optimum (30/22 °C) and low (22/14 °C) temperature conditions under optimum water and nutrient conditions. In Experiment II, plants were grown at optimum water (soil moisture content of 0.167 m³ m^?3) and in drought (soil moisture content 0.105 m³ m^?3) conditions under optimum temperature conditions. Above- and below-ground growth traits including several root traits of the CS lines were assessed at 25 days after sowing. The findings suggest which substituted chromosome or chromosome segment from the alien species likely harbors one or more genes for higher and lower tolerance to low temperature, respectively. CS-T04 and CSB08sh showed higher and lower tolerance to low temperature, respectively and CS-T04 and CS-B22sh showed higher and lower tolerance, respectively, to drought. CS lines are valuable analytical tool and useful genetic resources for targeted exploitation of beneficial genes for drought and low temperature stresses in Upland cotton.     

Development of InDel markers for the restorer gene <Emphasis Type="Italic">Rf1</Emphasis> and assessment of their utility for marker-assisted selection in cotton

The cytoplasmic male sterility (CMS) system is ideal for exploiting heterosis in crops such as cotton. However, CMS-D2, which is based on Gossypium harknessii cytoplasm, is still not widely used for cotton production. In this study, we developed an efficient marker-assisted selection method based on insertion/deletion (InDel) markers that can identify restorer lines carrying Rf1. Whole-genome resequencing was first completed for restorer [N(Rf1Rf1)] and maintainer [N(rf1rf1)] lines with normal fertile cytoplasm (N). Comparisons with the TM-1 reference genome sequence resulted in the identification of 292,065 and 183,657 InDels for the restorer and maintainer lines, respectively. Most of the InDels in the restorer line were distributed on Chromosome_D05, which carries Rf1. Of the 12 InDel markers near the Rf1 target region that were further validated, four co-dominant markers (i.e., InDel-1891, InDel-3434, InDel-7525, and InDel-9356) co-segregated with Rf1, as verified by a segregation analysis in an F₂ population. We subsequently used InDel-1891 to determine the allele status at the Rf1 locus in a backcross scheme for transferring Rf1. In this study, we developed new markers to increase the marker density in the Rf1 target region, which will be useful for the fine mapping of Rf1. The development of convenient and inexpensive co-segregating InDel markers will facilitate the marker-assisted selection of restorer lines carrying Rf1.     

Chromosomal location of genes for resistance to powdery mildew in <Emphasis Type="Italic">Agropyron cristatum</Emphasis> and mapping of conserved orthologous set molecular markers

Agropyron cristatum exhibits resistance to Blumeria graminis f. sp. tritici. Disomic and ditelosomic chromosome addition lines of A. cristatum in ‘Chinese Spring’ wheat were utilized to determine which A. cristatum chromosomes carry resistance gene(s). Resistance is conferred by gene(s) on chromosome arms 2PL and 6PL. The availability of molecular markers capable of detecting these chromosome arms in a wheat background would be very useful for marker-assisted introgression of 2PL and 6PL chromatin into common wheat. With this aim, 170 wheat conserved orthologous set (COS) markers (92 and 78 from wheat homoeologous groups 2 and 6 respectively) were assessed for their utility in A. cristatum. A total of 116 (68.2%) COS markers successfully amplified product in A. cristatum and 46 (40.0%) of these markers were polymorphic between A. cristatum and common wheat. From marker loci mapping on wheat homoeologous group 2 chromosomes, 23 markers (34.9%) were polymorphic between A. cristatum and common wheat and from them 13 markers were assigned to chromosome arm 2PL and six markers were mapped to chromosome 4P of A. cristatum showing that this chromosome is related to wheat homoeologous group 2. From marker loci mapping on wheat homoeologous group 6 chromosomes, 23 (46.0%) markers were polymorphic between A. cristatum and common wheat and from them 17 markers were located on chromosome 6P, six of them were mapped to chromosome arm 6PS and five to chromosome arm 6PL, respectively. The specific COS markers allocated on the long arms of chromosomes 2P and 6P may have a role in marker-assisted screening in wheat breeding for powdery mildew disease resistance.     

Overcoming obstacles to interspecific hybridization between <Emphasis Type="Italic">Gossypium hirsutum</Emphasis> and <Emphasis Type="Italic">G. turneri</Emphasis>

Gossypium turneri, a wild cotton species (2n = 2X = 26, D₁₀D₁₀) originating from Mexico, possesses invaluable characteristics unavailable in the cultivated tetraploid cotton gene pool, such as caducous involucels at anthesis, resistance to insects and tolerance to abiotic stresses. However, transferring desired characteristics from wild species into cultivated cotton is often fraught with diverse obstacles. Here, Gossypium hirsutum (as the maternal parent) and G. turneri were crossed in the Hainan Province of China, and the obtained hybrid seeds (2n = 3X = 39, ADD₁₀) were treated with 0.075% colchicine solution for 48 h to double the chromosome complement in order to overcome triploid F₁ sterility and to generate a fertile hexaploid. Chromosome doubling was successful in four individuals. However, the new synthetic hexaploids derived from these individuals were still highly sterile, and no seeds were generated by selfing or crossing. Therefore, an embryo rescue technique was employed in an attempt to produce progenies from the new synthetic hexaploids. Consequently, a total of six large embryos were obtained on MSB2K medium supplemented with 0.5 mg l^?1 KIN and 250 mg l^?1 CH using ovules from backcrossing that were 3 days post-anthesis. Four grafted surviving seedlings were confirmed to be the progenies (pentaploids) of the new synthetic hexaploids using cytological observations and molecular markers. Eight putative fertile individuals derived from backcrossing the above pentaploids were confirmed using SSR markers and generated an abundance of normal seeds. This research lays a foundation for transferring desirable characteristics from G. turneri into upland cotton.     

Molecular markers for improving control of soil-borne pathogen <Emphasis Type="Italic">Fusarium oxysporum</Emphasis> in sugar beet

Fusarium spp. cause severe damage in many agricultural crops, including sugar beet, with Fusarium oxysporum historically being considered as the most damaging of all species. Sugar beet needs to be protected from this class of soil-borne pathogens in order to ensure an optimal sugar yield in the field. Genetic control of the disease is crucial in managing these pathogens. Identification of single nucleotide polymorphism (SNP) markers linked to resistance can be a powerful tool for the introgression of valuable genes needed to develop Fusarium-resistant varieties. A candidate gene approach was carried out to identify SNP markers linked to putative Fusarium resistance sources in sugar beet. Five resistant analogue genes (RGAs) were screened by means of high resolution melting (HRM) analysis in a set of sugar beet lines, considered as resistant and susceptible to Fusarium oxysporum. HRM polymorphisms were observed in 80% of amplicons. Two HRM polymorphisms were significantly associated with Fusarium resistance (P < 0.05). The amplicons that showed association were sequenced and two SNPs were identified. The association was further validated on 96 susceptible and 96 resistant plants using competitive allele-specific PCR (KASPar) technology. The selected SNPs could be used for marker-assisted breeding of Fusarium resistance in sugar beet.     

Analysis of population structure and genetic diversity reveals gene flow and geographic patterns in cultivated rice (<Emphasis Type="Italic">O. sativa</Emphasis> and <Emphasis Type="Italic">O. glaberrima</Emphasis>) in West Africa

To fully exploit the diversity in African rice germplasm and to broaden the gene pool reliable information on the population genetic diversity and phenotypic characteristics is a prerequisite. In this paper, the population structure and genetic diversity of 42 cultivated African rice (Oryza spp.) accessions originating from West Africa (Benin, Mali and Nigeria, Liberia etc.) were investigated using 20 simple sequence repeats (SSR) and 77 amplified fragment length polymorphisms (AFLP). Additionally, field trials were set up to gain insight into phenotypic characteristics that differentiate the genetic populations among rice accessions. The analysis revealed considerably high polymorphisms for SSR markers (PIC mean?=?0.78) in the germplasm studied. A significant association was found between AFLP markers and geographic origin of rice accessions (R?=?0.72). Germplasm structure showed that Oryza sativa accessions were not totally isolated from Oryza glaberrima accessions. The results allowed identification of five O. glaberrima accessions which grouped together with O. sativa accessions, sharing common alleles of 18 loci out of the 20 SSR markers analyzed. Population structure analysis revealed existence of a gene flow between O. sativa and O. glaberrima rice accessions which can be used to combine several interesting traits in breeding programs. Further studies are needed to clarify the contributions of this gene flow to valuable traits such as abiotic and biotic stresses including disease resistance.     

Estimation of genetic diversity among 34 genotypes in the genus <Emphasis Type="Italic">Cajanus</Emphasis> with contrasting host response to the pod borer and its allied pests

The genetic divergence among 34 genotypes belonging to 12 species of genus Cajanus were carried out using plant pest interaction and DNA marker analysis. Principal component analysis based on average percentage of pod damage caused by pod borer, plume moth, and blue butterfly in the field conditions, and growth of their larva and pupa on an artificial diet in vitro dispersed these genotypes into four coordinates evincing high genetic divergence as expected. DNA marker analysis using 11 pairs of SSR and nine ISSR primers showed higher polymorphism at the species level, and these primers exhibited variation with regard to average band informativeness, resolving power, and PIC value. No single primer was able to distinguish between all the 34 genotypes of Cajanus but nine species specific amplified fragments were generated by five ISSR primers. The pairwise Jaccard’s similarity coefficient and Nei’s genetic distance values revealed a higher level of inter-specific genetic variation in the genus Cajanus. The clustering of genotypes based on Jaccard’s similarity coefficient vis-a-vis Nei’s genetic distance agreed with the sectional classification of the genus Cajanus. Seven cultivars of C. cajan and the genotypes of their wild progenitor C. cajanifolius remained in one cluster, whereas accessions of C. platycarpus and C. scarabaeoides were out grouped. The rest of the genotypes belonging to nine species of Cajanus formed another cluster. The principal coordinate analysis also supported this clustering pattern. Moreover, these findings have good many implications for future breeding endeavors aimed at the introgression of pod borer resistance alleles.     

Development of RAPD and ISSR derived SCAR markers linked to <Emphasis Type="Italic">Xca1Bo</Emphasis> gene conferring resistance to black rot disease in cauliflower (<Emphasis Type="Italic">Brassica oleracea</Emphasis> var. <Emphasis Type="Italic">botrytis</Emphasis> L.)

Black rot caused by Xanthomonas campestris pv. campestris (Xcc) (Pam.) is the most devastating disease of cauliflower (Brassica oleracea var. botrytis L.; 2n = 2x = 18), taking a heavy toll of the crop. In this study, a random amplified polymorphic DNA (RAPD) and inter simple sequence repeat (ISSR) derived sequence characterized amplified region (SCAR) markers linked to the black rot resistance locus Xca1bo were developed and evaluated as a screening tool for resistance. The RAPD marker OPO-04₈₃₃ and ISSR marker ISSR-11₆₃₅ were identified as closely linked at 1.6 cM distance to the black rot resistance locus Xca1bo. Both the markers OPO-04₈₃₃ and ISSR-11₆₃₅ were cloned, sequenced and converted into SCAR markers and validated in 17 cauliflower breeding lines having different genetic backgrounds. These SCAR markers (ScOPO-04₈₃₃ and ScPKPS-11₆₃₅) amplified common locus and showed 100% accuracy in differentiating resistant and susceptible plants of cauliflower breeding lines. The SCAR markers ScOPO-04₈₃₃ and ScPKPS-11₆₃₅ are the first genetic markers found to be linked to the black rot resistance locus Xca1bo in cauliflower. These markers will be very useful in black rot resistance marker assisted breeding.     

Further QTL mapping for yield component traits using introgression lines in rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.) under drought field environments

To better understand the underlying mechanisms of agronomic traits related to drought resistance and discover candidate genes or chromosome segments for drought-tolerant rice breeding, a fundamental introgression population, BC₃, derived from the backcross of local upland rice cv. Haogelao (donor parent) and super yield lowland rice cv. Shennong265 (recurrent parent) had been constructed before 2006. Previous quantitative trait locus (QTL) mapping results using 180 and 94 BC₃F_6,7 rice introgression lines (ILs) with 187 and 130 simple sequence repeat (SSR) markers for agronomy and physiology traits under drought in the field have been reported in 2009 and 2012, respectively. In this report, we conducted further QTL mapping for grain yield component traits under water-stressed (WS) and well-watered (WW) field conditions during 3 years (2012, 2013 and 2014). We used 62 SSR markers, 41 of which were newly screened, and 492 BC₄F_2,4 core lines derived from the fourth backcross between D123, an elite drought-tolerant IL (BC₃F₇), and Shennong265. Under WS conditions, a total of 19 QTLs were detected, all of which were associated with the new SSRs. Each QTL was only identified in 1 year and one site except for qPL-12-1 and qPL-5, which additively increased panicle length under drought stress. qPL-12-1 was detected in 2013 between new marker RM1337 and old marker RM3455 (34.39 cM) and was a major QTL with high reliability and 15.36% phenotypic variance. qPL-5 was a minor QTL detected in 2013 and 2014 between new marker RM5693 and old marker RM3476. Two QTLs for plant height (qPHL-3-1 and qPHP-12) were detected under both WS and WW conditions in 1 year and one site. qPHL-3-1, a major QTL from Shennong265 for decreasing plant height of leaf located on chromosome 3 between two new markers, explained 22.57% of phenotypic variation with high reliability under WS conditions. On the contrary, qPHP-12 was a minor QTL for increasing plant height of panicle from Haogelao on chromosome 12. Except for these two QTLs, all other 17 QTLs mapped under WS conditions were not mapped under WW conditions; thus, they were all related to drought tolerance. Thirteen QTLs mapped from Haogelao under WS conditions showed improved drought tolerance. However, a major QTL for delayed heading date from Shennong265, qDHD-12, enhanced drought tolerance, was located on chromosome 12 between new marker RM1337 and old marker RM3455 (11.11 cM), explained 21.84% of phenotypic variance and showed a negative additive effect (shortening delay days under WS compared with WW). Importantly, chromosome 12 was enriched with seven QTLs, five of which, including major qDHD-12, congregated near new marker RM1337. In addition, four of the seven QTLs improved drought resistance and were located between RM1337 and RM3455, including three minor QTLs from Haogelao for thousand kernel weight, tiller number and panicle length, respectively, and the major QTL qDHD-12 from Shennong265. These results strongly suggested that the newly screened RM1337 marker may be used for marker-assisted selection (MAS) in drought-tolerant rice breeding and that there is a pleiotropic gene or cluster of genes linked to drought tolerance. Another major QTL (qTKW-1-2) for increasing thousand kernel weight from Haogelao was also identified under WW conditions. These results are helpful for MAS in rice breeding and drought-resistant gene cloning.     

Obtainment of an intergeneric hybrid between <Emphasis Type="Italic">Forsythia</Emphasis> and <Emphasis Type="Italic">Abeliophyllum</Emphasis>

Forsythia suspensa and F. ‘Courtaneur’ were used as female parents to cross with Abeliophyllum distichum in 2011 and an intergeneric hybrid of F. suspensa × A. distichum was obtained, though with very low seed set. The morphological characteristics, flower fragrance and volatile organic compounds of flowers were analysed. The intergeneric hybrid had intermediate morphological characteristics of both parents and flower fragrance and was confirmed as a true intergeneric hybrid by amplified fragment length polymorphism (AFLP) markers. Compared with its mother parent (F. suspensa), flowers of the intergeneric hybrid are pale yellow with delicate fragrance. Volatile organic compounds of flowers were retrieved by purge-and-trap techniques, and determined by gas chromatography and mass spectrometry (GC–MS). The main volatile organic components of F. suspensa were isoprenoids, while the main volatile organic components of A. distichum and the hybrid of F. suspensa × A. distichum were aliphatics. To determine the time and the site of intergeneric hybridizing barriers occured, the pollen tubes’ behavior after pollination was observed under fluorescence microscopy. It was found that significant pre-fertilization incompatibility existed in intergeneric crossing combinations [F. ‘Courtaneur’ (Pin) × A. distichum (Thrum) and F. suspensa (Pin) × A. distichum (Thrum)], and only a few pollen tubes of A. distichum penetrated into the ovaries of Forsythia. In our research, an intergeneric hybrid between Forsythia and Abeliophyllum was obtained for the first time, which will provide a solid foundation for expanding the flower color range of Forsythia and breeding fragrant-flowered cultivars.     

<Emphasis Type="Italic">Er3</Emphasis> gene,conferring resistance to powdery mildew in pea,is located in pea LGIV

Three genes for resistance to Erysiphe pisi, named er1, er2 and Er3 have been described in pea so far. er1 gene is located in pea linkage group VI, while er2 gene has been mapped in LGIII. SCAR and RAPD markers tightly linked to Er3 gene have been identified, but the position of these markers in the pea genetic map was unknown. The objective of this study was to localize Er3 gene in the pea genetic map. Towards this aim, the susceptible pea cv. Messire (er3er3) and a resistant near isogenic line of Messire (cv. Eritreo, Er3Er3) were surveyed with SSRs with known position in the pea map. Three SSRs were polymorphic between “Messire” and “Eritreo” and further surveyed in two contrasting bulks formed by homozygous Er3Er3/er3er3 individuals obtained from a F₂ population derived from the cross C2 (Er3Er3)?×?Messire (er3er3). A single marker, AA349, was polymorphic between the bulks. Subsequently, other ten markers located in the surrounding of AA349 were selected and analysed in Er3Er3 and er3er3 plants. As a results, another SSR, AD61, was found to be polymorphic between Er3Er3 and er3er3 plants. Further linkage analysis confirmed that SSRs AA349 and AD61 were linked to Er3 and to the RAPD and SCAR markers previously reported to be linked to this gene. Er3 gene was located in pea LGIV at 0.39 cM downstream of marker AD61. The location of Er3 gene in the pea map is a first step toward the identification of this gene.     

Identification of SNP for rice blast resistance gene <Emphasis Type="Italic">Pike</Emphasis> and development of the gene-specific markers

Rice blast disease caused by Magnaporthe oryzae is an important limiting factor to rice production in the world. Introgression of blast resistance genes into improved germplasm by marker-assisted selection has been considered as an effective and environmentally beneficial means to control this disease. Pike, a broad-spectrum blast resistance gene, was cloned by map-based strategy recently in our laboratory. Two adjacent CC-NBS-LRR genes (designated as Pike-1 and Pike-2) were required for Pike-mediated resistance. In the current study, sequence alignment of the SNP G1328C and the SNP-surrounding region let us find that the Pik DNA variants of the studied rice lines appear to be divided into G-, C-, T- and G’-types. Based on the four genotypes, a Pike-specific marker system consisting of three PCR-based markers CP-G1328C, CP-G1328T and CP-G1328G’ was developed and used to effectively differentiate G-type allele from each of the others. Using this marker system, we investigated distribution of the Pik DNA variants in a set of 326 rice varieties or breeding lines and found that there were 2, 130, 135 and 59 rice lines identified to carry G-, C-, T- and G’-type alleles, respectively. In addition, with sequence data of the SNP G1328C-containing genomic region derived from 56 rice lines, we constructed a phylogenetic tree with three major clades which just corresponded to the types of the Pik DNA variants described above.     

Overcoming unilateral incompatibility in crosses with wild allotetraploid potato species <Emphasis Type="Italic">Solanum stoloniferum</Emphasis> Schldtl. &amp; Bouchet

The germplasm of valuable for breeding wild allotetraploid potato species Solanum stoloniferum is rarely used because of pre- and postzygotic reproductive barriers with cultivated potatoes. One of the factors that complicate crosses between S. stoloniferum and S. tuberosum is unilateral incompatibility (UI). Here, we present the results of application of S. verrucosum and S _v S _v -lines for overcoming UI in crosses with S. stoloniferum and of generating male fertile hybrids derived from this species. S _v S _v -lines are F2 S. tuberosum dihaploid × S. verrucosum that are male fertile and have D/γ-type cytoplasm. Since they are homozygous for S _v gene from S. verrucosum, they were expected to have the same ability for elimination of prezygotic incompatibility as this species. Three accessions of S. verrucosum and seven S _v S _v -lines were pollinated by 26 accessions of S. stoloniferum. The crosses with S. verrucosum failed or had low efficacy (1.5–2.4 seeds per pollination). On the other hand, use of S _v S _v -lines was more efficient: 15.8 seeds per pollination. In spite of low percentage of germination (1.9%), 40 seedlings of interspecific hybrids were produced. The experiment on hybridization between S _v S _v -lines and S. stoloniferum has been reproduced with the accession PI 205522 of the wild species, which had DNA markers of PVY and LB resistance genes and W/γ cytoplasm: 950 hybrid seeds and 12 viable seedlings were produced. The genome of the seedlings was doubled by colchicine treatment, which generated hexaploids that formed highly fertile pollen and set seeds from self-pollination. We were able to cross them as females with the variety Katahdin.     

Exploring ethyl methanesulfonate (EMS) treated cotton (<Emphasis Type="Italic">Gossypium hirsutum</Emphasis> L.) to improve drought tolerance

The Texas High Plains often has extended periods between rainfall events, which can lead to a reduction in the yield and fiber quality of cotton (Gossypium hirsutum L.). It is known that cultivated cotton suffers from low levels of genetic diversity due to the over-use in breeding of similar gene pools, which may hinder breeding for drought tolerance. In this study, for the first time the novel variability or genetic diversity of morphological and agronomic traits possibly created by the chemical mutagen ethyl methanesulfonate (EMS) was evaluated to improve drought tolerance in cotton by traits’ response to different irrigation regimes. EMS is a chemical mutagen that has been shown to cause point mutations in the DNA of many model plants and crop species. Three EMS treated lines were advanced from the M₁ to M₄ generation as bulk-harvested populations. A diverse selection scheme was applied to capture most of the genetic trait-variability or diversity and superior lines in these populations. In 2014–2016 the diversity of these populations was evaluated based on four agronomic and thirteen morphological traits to determine differences in response to multiple irrigation rates. Analyses of these traits showed statistically significant (p?≤?0.05) differences between and within populations when compared to the original non-treated EMS source, with most of the variability being observed in the high irrigation rate. However, none of the EMS treated populations had significantly (p?≤?0.05) better lint yield than the commercial cultivar (control) in 2016. EMS yield performance was possibly constrained by the applied diverse selection scheme of this study. Traits such as total number of bolls, bolls retained at node 7 and below, and those retained between nodes 8 and 12, and bolls retained at node of first fruiting branch may be predictors to improve cotton production (yield) in water limiting environments.