Development of AFLP and derived CAPS markers for root-knot nematode resistance in cotton

Resistance to root-knot nematode (Meloidogyne incognita) is determined by a single major gene rkn1 in Gossypium hirsutum Acala NemX cotton. Bulked segregant analysis (BSA) combined with amplified fragment length polymorphism (AFLP) was used to identify molecular markers linked to rkn1. DNA pools from homozygous susceptible (S) and resistant (R) bulks of an F_2:3 originating from the intraspecific cross NemX × SJ-2 were screened with 128 EcoR1/Mse1 primer combinations. Putative AFLP markers were then screened with 60 F_2:7 RIL plants and four AFLP markers were found linked to rkn1. The linkage of AFLP markers to rkn1 was also confirmed in a F₂ population. The closest AFLP marker was converted to a cleaved amplified polymorphic sequence (CAPS) marker (designated GHACC1) by aligning the sequences from both susceptible and resistant parents. GHACC1 linkage to rkn1 was confirmed in the F₂ (1R:3S), F_2:7 RIL (1R:1S) and the backcross population SJ-2 × F₁ (NemX × SJ-2) (1 heterozygous: 1 homozygous). The four AFLP markers, GHACC1 plus two SSR markers (CIR316 and BNL1231) linked to rkn1 from previous work were mapped to intervals of 2.6–14.2 cM from the rkn1 locus, and the genomic region around rkn1 was spanned to about 28.2 cM in the F_2:7 population. The PCR-based GHACC1 and CIR316 markers were tested on 21 nematode resistant and susceptible cotton breeding lines and cultivars. GHACC1 was suitable for nematode resistance screening within G.␣hirsutum, but not G. barbadense, whereas CIR316 was useful in both species, indicating their␣potential for utilization in marker-assisted selection.     

油菜素内酯在棉花组织培养中的应用研究Ⅰ．油菜素内酯对陆地棉体细胞胚胎发生和根器官发生的影响

本文研究了ＢＲ及其与ＩＡＡ、２，４－Ｄ、ＫＴ和６－ＢＡ等配合使用对陆地棉愈伤组织诱导、继代、分化、体细胞胚胎和根器官发生的影响。ＢＲ０．０１ｍｇ／Ｌ能使陆地棉Ｃｏｋｅｒ２０１、３１２两品种分化产生胚性愈伤组织，并有效地保持该种愈伤组织的生活力和胚胎发生能力。ＢＲ０．０１ｍｇ／Ｌ＋ＩＡＡ０．５ｍｇ／Ｌ促进Ｃｏｋｅｒ２０１、３１２两品种体细胞胚胎发生，ＢＲ０．０１ｍｇ／Ｌ＋２，４－Ｄ０．０５ｍｇ／Ｌ能诱导所有供试品种产生疏松黄绿色愈伤组织，２，４－Ｄ用量逐步降低或除去后，一些品种便分化产生胚性愈伤组织或体细胞胚状体。另在ＢＲ与ＩＡＡ的某些组合中还观察到根器官的发生。     

Genetic analysis of in vitro wheat somatic embryogenesis

Summary A spring wheat genotype which produces somatic embryos in vitro, after short and long-term culture, was tested for its ability to sexually transmit this embryogenic trait. Reciprocal crosses were performed between a embryogenic line and a nonembryogenic variety.Immature embryos were cultured on Murashige and Skoog medium plus 2 mg/l 2,4-dichlorophenoxyacetic acid, gelled with 5.5 g/l agarose. Somatic embryogenesis was not expressed in the F₁'s. In contrast, from several hundred immature embryos of the F₂ generation of one cross, 10.7% and 1.6% expressed somatic embryogenesis in short and long-term cultures respectively. These percentages of embryogenic: non-embryogenic fits a model of a few complementary genes. The embryogenic capacity of the F₂ genotypes depends on the presence of recessive alleles at these gene loci. The long-term wheat somatic embryogenesis capacity requires a more complex mechanism than the short-term one.Abbreviations CS Chinese Spring - Aq Aquila - E Embryogenic - NE Nonembryogenic - SC Subculture     

Rhizobium specificity of Trifolium ambiguum M. Bieb × T. repens L. hybrid clovers

Growth of full-sib families of an F₁ interspecific hybrid between Trifolium ambiguum M.Bieb and T. repens L, and two generations of backcross hybrids (BC₁F₂ and BC₂F₁) with T. repens as the recurrent parent, were compared to their parental species. Plants were grown in a N-free medium and inoculated with Rhizobium leguminosarum biovar trifolii rhizobia effective on T. ambiguum or T. repens. Hybridisation produced progeny that nodulated with rhizobia from either T. ambiguum or T. repens, but plant growth varied. Mean weights of T. repens and hybrids, particularly F₁ and BC₁F₂, were higher when inoculated with a mixture of strains isolated from field grown T. repens than with the New Zealand inoculant strain for T. repens. When inoculated with the mix of rhizobia from T. repens, mean weights were 242, 189, 132, 125, and 100 mg/plant for T. repens, BC₂F₁, BC₁F₂, F₁ and T. ambiguum, respectively. However, although the mean weight of BC₂F₁ lines was significantly less than T. repens, there was considerable variation in individual full-sib families indicating the potential to select within BC₂F₁ hybrids for high plant growth/symbiotic nitrogen fixation. The weight of T. ambiguumplants inoculated with the New Zealand inoculant strain for hexaploid T. ambiguum was similar to T. repens inoculated with the mix of rhizobia from T. repens (253 and 242 mg/plant, respectively). Mean fresh weights of F₁ hybrid plants were similar when inoculated with rhizobia for T. ambiguum or T. repens (125 and 130 mg/plant, respectively). However, weight of T. repens, BC₁F₂ and BC₂F₁ hybrids inoculated with rhizobia for T. ambiguum were all less than 90 mg/plant. There was a significant relationship between plant fresh weight and ethylene production. The results indicate that measuring weights of inoculated plants growing in N-free media is a rapid initial method of screening a range of plant germplasm for plant growth/symbiotic nitrogen fixation rates. This revised version was published online in August 2006 with corrections to the Cover Date.     

Identifying QTL for fiber quality traits with three upland cotton (Gossypium hirsutum L.) populations

Cotton fiber quality was quantitative trait, controlled by multiple genes. Identification of stable quantitative trait loci (QTL) effectively contributing to favorable fiber quality traits would provide the key basis for marker-assisted selection used in molecular breeding projects. Three upland cotton F₂ populations were established with a common parent Chinese cultivar Yumian 1 and three American commercial cultivars/lines (Acala Maxxa, CA3084 and TAM94L-25), each of which had unique fiber quality characteristic that was favorable economically. Three whole genome genetic maps were constructed with 323, 302 and 262 SSR loci for population (Yumian 1 × Acala Maxxa), (Yumian 1 × CA3084), and (Yumian 1 × TAM 94L-25) respectively, spanning 1,617.2, 1,639.9 and 1,441.4 cM. Based on these genetic maps and three generation phenotypic data of fiber quality traits (F₂, F_2:3 and F_2:4), 77 QTL were detected, including 19 for fiber length, 14 for fiber uniformity, 17 for micronaire, 10 for fiber elongation, and 17 for fiber strength. Among these QTL, 46 QTL were significant QTL and 31 were putative QTL, including that one QTL (qFL05.1) and four QTL (qFL23.1, qFM06.1, qFM06.2 and qFE25.1) were detected across three and two populations respectively; two QTL qFL10.1 (Yumian 1 × TAM 94L-25) and qFL15.1 (Yumian 1 × Acala Maxxa) were detected in three generations; qFM23.1, qFE18.1 and qFS21.2 detected in population (Yumian 1 × CA3084), qFE10.1, and qFS10.2 detected in population (Yumian 1 × TAM 94L-25), and qFS15.1 detected in population (Yumian 1 × Acala Maxxa), were all detected in two generations. Alleles underlying these stable QTL were valuable candidate gene for fine mapping, cloning, and favorable gene pyramiding projects. Our study also verified that QTL mapping of fiber quality traits using multiple populations with a common parent had higher efficiency compared to single population crossed with two parents and favorable alleles contributed to QTL effect could be conferred by parents with inferior fiber quality traits.     

Sexual Transmission of Somatic Embryogenesis in Dactylis glomerate*

A Dactylis glomerata L. genotype that produces somatic embryos in vitro was tested for the ability to sexually transmit the embryogenic trait. Reciprocal crosses were performed between the embryogenic and two non-embryogenic genotypes. Leaf segments from 69 F₁ plants were cultured on Schenk and Hildebrandt medium amended with 30 μM 3,6-dichloro-o-anisic acid (dicamba). Somatic embryogenesis was expressed in 39 of the F₁ plants. The embryogenic parent was female for 18 of these plants and male for the other 21. The 39:30 ratio of embryogenic: non-embryogenic fits an expected 1:1 for tetrasomic inheritance of a dominant nuclear gene.     

Genetic control of somatic embryogenesis in alfalfa (Medicago sativa L. cv. Adriana)

Summary In alfalfa (Medicago sativa) regeneration is genotype-specific. In order to study the genetic control of somatic embryogenesis and to constitute a synthetic cultivar characterized by its high regeneration ability, 2 embryogenic plants selected from the cv. Adriana were selfed, intercrossed and also crossed in both directions with 5 non-embryogenic genotypes of the same cultivar.Progenies of all crosses were scored for their regeneration ability and results indicate that somatic embryogenesis is under the control of 2 dominant loci. However some non-embryogenic genotypes prevent regeneration when crossed with embryogenic ones and this characteristic is not under the control of a single dominant gene.When plants chosen for their capacity to regenerate within F₁ and S₁ progenies were freely intercrossed the regeneration efficiency dropped to 2% (1 plant out of 50). This result indicates that if the genetic background of the population is changed the regeneration is greatly affected and therefore some other mechanism could play a role in determining plant regeneration.     

Identification and inheritance of male sterility in groundnut (Arachis hypogaea L.)

Summary Some plants without pods but with gynophores were observed in two F₄ progenies of two crosses of goundnut (Arachis hypogaea L.). The flowers on these plants had translucent white anthers with no or a few sterile pollen grains. Three such plants in the succeeding generation were hand pollinated with pollen from a short-duration Indian cv. JL 24. The resulting F₁ hybrid plants (male sterile x JL 24) were normal. Chi-square tests for segregation for male fertile and male sterile plants in F₂ and F₃ generations indicated that the male sterility in these crosses of groundnut is governed by two recessive genes. We designate these genes as ms₁ and ms₂ with ms₁ms₁ms₂ms₂ being a male sterile genotype.Submitted as ICRISAT J. A. No. 1812.     

RAPD markers linked to microspore embryogenic ability in Brassica crops

In order to identify the markers linked to microspore embryogenic ability in Brassica crops, RAPD segregation analyses were performed in a microspore-derived (MD) population and a F₂ population derived from F₁between ‘Ho Mei’ (high responsive parent in microspore embryogenesis) and ‘269’ (low responsive parent) in Chinese cabbage, and between ‘Lisandra’ (high responsive parent) and ‘Kamikita’ (low responsive parent) in oil seed rape. After 230 and 143 primers were screened, a total of 148 and 52markers were detected to be polymorphic between the parents in Chinese cabbage and oilseed rape, respectively. Twenty-seven percent of the markers in the MD population showed a significant segregation distortion in both crops. Of the markers showing segregation distortion in the MD population, 71–75% of the markers followed the expected Mendelian segregation ratio in the F₂ population. When the relationships between such distorted markers and microspore embryogenesis of the F₂ population were examined, 7 and 3 markers were identified to be associated with embryogenic ability in Chinese cabbage and oilseed rape, respectively. These markers showed additive effects on embryo yields, and the plants having more alleles of the high responsive parent produced higher embryo yields. These markers maybe useful in marker-assisted selection for improving microspore responsiveness straits in Brassica crops. This revised version was published online in July 2006 with corrections to the Cover Date.     

Monosomic analysis of stem rust resistance in the wheat cultivar Almus

Summary Monosomic analysis of resistance to stem rust, race 11 (isolate G 425) was carried out in the cultivar Almus (GDR) possessing a 1B/1R translocation. F₂ progenies of monosomic and disomic F₁ plants of Almus crossed with 21 monosomic lines of Chinese Spring were tested. Two lines (1B and 6B) differed significantly from the disomic segregation ratio by a higher number of resistant plants and two other lines (1D and 6A) by a lower number of resistant plants. The results fitted a hypothesis comprising the interaction of two genes for resistance and two inhibitors.     

Increasing embryogenesis and doubling efficiency by immediate colchicine treatment of isolated microspores in spring Brassica napus

The effect of colchicine on induction of embryogenesis andchromosome doubling during microspore culture was evaluated in twoF₁ hybrids of spring oilseed rape (Brassica napus L.). Immediatecolchicine treatment of isolated microspores with the concentrations 50 and500 mg/L for 15 h stimulated embryogenesis and produced largeamounts of healthy-looking embryos. These normal embryos germinatedwell at 24 ^°C after being transferred to solid regeneration mediumand an initial period of low temperature (2 ^°C) for 10 days, andcould directly and rapidly regenerate vigorous plants. A high doublingefficiency of 83–91% was obtained from 500 mg/L colchicinetreatment for 15 h with low frequency of polyploid and chimeric plants.The present experiment showed that a treatment duration of 30 h revealedless positive effects on embryogenesis and doubling efficiency, especially athigher colchicine concentration (1000 mg/L). Poor embryogenesis andembryo germination were observed from ordinary microspore culturewithout change of induction medium and colchicine treatment, and severalsubcultures were required for induction of secondary embryogenesis andplant regeneration.     

Genetic variance components and genetic effects among eleven diverse upland cotton lines and their F2 hybrids

Selecting high yielding upland cotton, Gossypium hirsutum L. lines with improved fiber quality is a primary breeding goal. A diverse set of ten cultivars and one breeding line were crossed in a half diallel. Parents and F₂ hybrids were grown in three environments at Mississippi State, MS. Ten agronomic and fiber traits were analyzed by a mixed linear model approach based on the additive-dominance genetic model. Variance component, genetic effects and genetic correlations were calculated. ‘Acala Ultima’ was a desirable general combiner for fiber length, uniformity, strength, micronaire, lint percentage, and boll weight. ‘FiberMax 966’ was a desirable general combiner for fiber length, uniformity, strength, and all agronomic traits. ‘Tamcot Pyramid’ and M240 were poor general combiners for both fiber and agronomic traits. ‘Coker 315’ was a good general combiner for fiber length, uniformity, micronaire, boll weight, boll number, and yield. Heterozygous dominance effects were associated with several crosses, which suggest their use as hybrids.     

Evidence for the production of unreduced gametes by tetraploid Rosa hybrida L.

The objective of this study was to determine the mode of inheritance of field resistance to downy mildew (Peronospora parasitica (Pers. ex Fr.) Fr.) in broccoli (Brassica oleracea var.italica) at the adult plant stage. The F₁, F₂ and F₃ progeny of resistant and susceptible plants of broccoli were tested in the field under natural infection, in central Portugal, from August to December in two successive years. The plants were evaluated for resistance to downy mildew at maturity using a five-class scale of increasing susceptibility to the disease, which took into account the number of infected leaves and the size of the sporulating lesions. The F₁ was completely resistant, the F₂ segregated a clear 3 resistant: 1susceptible and the F₃ confirmed the F₂ segregation, which suggests a dominant character controlled by a single locus. This resistance has good potencial for direct use in commercial broccoli breeding or for transfer to other Brassica vegetables. This revised version was published online in August 2006 with corrections to the Cover Date.     

Crossability and cytology of hybrid progenies in the cross between Brassica campestris and three wild relatives of B. oleracea,B. bourgeaui,B. cretica and B. montana

Summary Crossability and cytology were examined in F₁, F₂, B₁ and hybridsplants of F₁ hybrids of Brassica campestris and three wild relatives of B. oleracea, B. bourgeaui, B. cretica and B. montana, respectively. The F₂ plants were obtained after self-and open pollination of the F₁ hybrids. The B₁ and hybrid plants were produced after the F₁ hybrids backcrosses with B. campestris and crossed with B. napus, respectively. After crossing the F₁ hybrids, many seeds of the F₂, B₁ and hybrid plants were harvested. Multivalent formation was high in the chromsome configuration for the PMCs of F₂, B₁ and hybrid plants, suggesting that crossing over might occur between them. Many different types of aneuploids were obtained in the progenies of the F₂, B₁ and hybrid plants. It is suggested that different types of normal egg cells may be produced by one-by-one or little-by-little chromosome addition. The possibility is discussed of gene transfer from B. bourgeaui, B. cretica and B. montana, to cultivated plants, B. campestris and B. napus.     

Research on the inheritance of fasciation in lettuce (Lactuca sativa L.)

Summary Inheritance of fasciation was investigated in reciprocal crosses between two lettuce varieties: Suzan (non-fasciated) and Noran (fasciated). In glasshouses parental plants, F₁, F₂, and F₃ populations were assessed for fasciation according to a scale from 0 (non-fasciated)-9 (extremely fasciated).The observed environmental variation for fasciation of the homozygous parents was very low or absent, but the environmental variation for the F₁ populations was large. On the basis of this large variation of the heterozygous F₁ plants we supposed that such a variation also occurred in F₂ and F₃ populations.Considerable mortality was observed in all generations, which supposedly occurred at random in parents and F₁ but was non-random in the segregating F₂ and F₃ populations. So, means and variances of F₂ populations will be biased and therefore they were not used for genetical analyses of fasciation.From F₁ and F₃ populations and from some F₂ populations it appeared that no differences occurred between reciprocals. The differences for fasciation between reciprocal F₂ populations in the 1977 A experiment may result from non-random plant survival. A regression of F₃-means on the values for fasciation of parental F₂ plants, adjusted for their inbreeding, resulted in a realized h²-narrow of 0.4. This indicates that in the surviving plants of the F₂ populations still additive genetic variation was present to select successfully for non-fasciated plants.     

Genetics of self-compatibility in dihaploids of Solanum tuberosum L. I. Breeding behaviour of two self-compatible dihaploids

Summary Two highly fertile and self-compatible dihaploids (2ns=2x24) from Solanum tuberosum L. (2n 4x 48) were investigated to elucidate the genetic basis of their self-compatibility. To this end the two dihaploids were selfed and reciprocally intercrossed and the resulting I₁ and F₁ plants tested for self-compatibility. Reciprocal backcrosses of I₁-plants and F₁-plants were made. Complete diallels both within self-compatible and within self-incompatible F₁-plants were carried out as well as reciprocal matings between self-compatible and self-incompatible F₁-plants. From the wealth of data it could be concluded, that the dihaploids have two intact S-alleles, one being common to both. Six hypotheses were tested for explaining self-compatibility in these particular dihaploids. All but one had to be discarded. It is concluded that the self-compatibility most likely is brought about by the presence of an S-bearing translocation, which is not linked to the S-locus. The ratio sc :si in the F₁'s point either to certative disadvantage of translocation-bearing pollen or to lethality of translocation homozygotes. The importance of this self-compatibility mechanism for genetic and breeding research in potato is discussed.     

Resistance to the cabbage root fly, Delia radicum, within Brassica fruticulosa

Two transgenic Bt rice lines, KMD1 and KMD2, both containing a synthetic cry1Ab gene from Bt, were crossed with conventional rice varieties. The inheritance of resistance to SSB of KMD1 and KMD2was investigated through LSB and field examination of their progenies, e.g. F₁, BC₁ and F₂ populations. In LSBs, 100.0% of newly hatched SSB larvae died on the second day after feeding on leaf tissues of F₁ and GUS positive BC₁ plants, of which the area of leaf tissues consumed by SSB is also similar to that of transgenic parents. These results imply that the resistance of Bt rice to SSB is dominantly controlled and could be easily exploited in hybrid rice production. Field evaluation showed that segregation ratios for SSB resistance to susceptibility in BC₁ populations fit the ratio of 1:1, which was also confirmed by LSBs. However, in F₂ populations, the ratio was significantly smaller than 3:1 for resistant to susceptible plants in all 6 indica × japonica (KMD1 and KMD2) crosses, though it fitted 3:1 in all 4 japonica × japonica crosses. The results implied that the resistance of Bt rice to SSB was controlled by a dominant gene which was present in a homozygous condition in both KMD1 and KMD2, but the inheritance could be affected by other factors. Assays for Cry1Ab protein showed that, in most crosses, the content of Cry1Ab is significantly higher in leaves of GUS positive F₁, BC₁ and F₂ plants than that in transgenic Bt parent plants, which accounts for the high resistance observed in these plants to SSB. This revised version was published online in July 2006 with corrections to the Cover Date.     

Chloroplast-DNA variation in the wild and cultivated olives (Olea europaea L.) of Morocco

The male sterile plants that segregated in a BC₅F₂ of `C. sericeus × C. cajan var. TT-5' population were maintained by sib mating. The male sterile plants were crossed with ICPL-85012.Approximately 50% of the F₁ plants were sterile. F₂ plants derived from the fertile F₁ plants did not segregate for male sterility. The reciprocal hybrid i.e. ICPL-85012 × Fertile derivatives from C. sericeus × TT-5, did not express male sterility. However, among the 12 F₂ plant to row progenies, two segregated 25% male sterile plants and remaining 10 did not segregate. The segregation pattern in subsequent progenies revealed that the sterility was under control of a single recessive allele. Studies on the backcross and their BC₁F₂ and BC₁F₃progenies revealed another sterility gene which was found to be dominant in inheritance. This paper shows that what was thought to be cytoplasmic male sterility from C. sericeus cytoplasm is actually a single dominant gene possibly acting in concert with a single recessive gene to mimic cytoplasmic male sterility. This revised version was published online in July 2006 with corrections to the Cover Date.     

Haploid induction from F1 hybrids between CMS shallot with Allium galanthum cytoplasm and common onion by unpollinated flower culture

The induction of haploid plants from F₁ hybrids between CMS shallot with Allium galanthum cytoplasm and common onion was examined. Starting with 535 unpollinated flowers cultured in B5 medium 25 seedlings from part henogenetic embryos were obtained of which 13 seedlings survived. Eleven seedlings were determined as haploid plants (2n = x = 8) and 2 seedlings were doubled haploid plants (2n = 2x = 16). All haploid and doubled haploid plants preserved chloroplast DNA (cpDNA) and mitochondrial DNA (mtDNA) from A. galanthum. Segregation in different characters was observed among the haploid plants. The haploid and doubled haploid plants exhibited the different combinations of genes from shallot and common onion. Crossing of the doubled haploid plants with other shallot strains, common onion cultivars or related species may produce excellent F₁ hybrids for bulb production. This revised version was published online in July 2006 with corrections to the Cover Date.     

Dominance of bush growth habit in spaghetti squash (Cucurbita pepo)

Summary Bush and vine habit accessions and their reciprocal F₁s of spaghetti squash (Cucurbita pepo) were compared for various vegetative and reproductive characteristics. Bush habit was dominant to vine. Vine plants produced more leaves, longer internodes, and more male flowers than did bush and F₁ plants. Pistillate flowers developed at lower nodes in bush and F₁ plants than in vine plants. Vine plants produced as much or more vegetative matter than did bush and F₁ plants, but bush and F₁ plants produced more fruits and greater yields than did vine plants.