Cytogenetics of a new trispecies hybrid in cotton: [(Gossypium hirsutum L. × G. thurberi Tod.)2 × G. longicalyx Hutch. & Lee]

A three‐species hybrid named HTL including Gossypium hirsutum L. [2n = 4 x = 52, (AD)₁ genome] was created using the pseudophyletic introgression method with G. longicalyx Hutch. & Lee (2n = 2x = 26, F₁ genome) as donor parent and G. thurberi Tod. (2n = 2x = 26, D₁ genome) as bridge species. The new hybrid was totally self‐sterile and its interspecific status was confirmed using simple sequence repeat markers and cytogenetic analysis. Cytogenetic studies showed that its chromosome configuration was 2n = 52 = 14.13 I + 15.10 II + 1.03 III + 0.9 IV + 0.03 V + 0.13 VI (where I, II, III, IV, V and VI are univalents, bivalents, trivalents, tetravalents, pentavalents and hexavalents, respectively). Prospects for successfully exploiting the HTL hybrid in breeding programmes are discussed.     

Validation of a set of informative simple sequence repeats markers for variety identification in Pak‐choi (Brassica rapa L. ssp. chinensis var. communis)

A core set of 21 simple sequence repeats (SSR) markers was developed for Pak‐choi (Brassica rapa ssp. chinensis var. communis) variety identification. We initially selected 74 SSR markers which exhibited high polymorphism and reproducibility in SSR detection from 2129 SSRs. Using the 74 SSR‐based dendrogram for 45 inbred lines as calibration, 21 core SSRs were selected out. The utility of this core set SSRs was firstly tested in 45 inbred lines and finally verified in 102 commercial varieties. We also constructed a molecular ladder for each core SSR as a reference standard. Diversity analysis of this core SSR panel in 102 varieties demonstrated that each marker generates 2–3 alleles (averaged 2.33), with polymorphism information content values ranging from 0.01 to 0.56 (averaged 0.31). The averaged values of Shannon information index, observed heterozygosity, expected heterozygosity and Wright's fixation index were 0.59, 0.43, 0.38 and −0.09, respectively. Furthermore, the 21 SSR‐based classifications for 102 varieties were consistent with traditional classification based on morphology. This core SSR panel represents an effective tool for genetic variation analysis in Pak‐choi.     

The resistance to leaf rust and powdery mildew of recombinant lines of barley (Hordeum vulgare L.) derived from H. vulgare × H. bulbosum crosses

A set of 23 recombinant lines (RLs) of barley ( Hordeum vulgare L.) derived from H. vulgare  ×  H. bulbosum L. crosses was inoculated with barley leaf rust ( Puccinia hordei ) and powdery mildew ( Blumeria graminis f.sp. hordei ) at the seedling stage to identify their levels and mechanisms of resistance. Eight RLs were studied further in glasshouse and field tests. All three barley parents ('Emir', 'Golden Promise' and 'Vada') were highly susceptible to powdery mildew and leaf rust isolates. Several RLs showed partial resistance expressed as high relative latency periods and low relative infection frequencies against leaf rust. This high level of partial resistance was due to a very high level of early aborting colonies without host cell necrosis. Several RLs showed hypersensitive resistance to some or all isolates. For powdery mildew, one RL was completely resistant to the CC1 isolate and had a hypersensitive resistance to the CO-02 isolate. Three RLs derived from 'Emir' were completely resistant to both powdery mildew isolates, and three more RLs tested in the field had higher levels of partial resistance than their parents. The results indicate that H. bulbosum contains major and minor gene(s) for resistance to leaf rust and powdery mildew that can be transferred to cultivated barley.     

Development of drought‐tolerant breeding lines derived from Helianthus annuus × H. argophyllus interspecific crosses

Helianthus argophyllus is a wild species known as “silver sunflower”, which possesses several traits, including morphological traits that increase drought tolerance. Therefore, introgressions between chosen cultivated lines and two H. argophyllus accessions were made, and segregating generations were established. Important agronomic traits including single heading, high pollen fertility, silver canopy (indicating more cuticular wax), and content of cuticular waxes and oil were selected over six segregating generations. The resulting F₆ lines showed introgression of water saving traits, as they had lower excise leaf water loss, with comparable yield to standard checks. However, these F₆ lines were late maturing and showed poor flowering synchronization between the cultivated and introgressed lines. Introgressed line “D‐22” was particularly promising as a breeding line, with superior agronomic and drought resistance traits. This line had the potential to be used as an inbred parental line for introgression of drought resistance traits into elite sunflower germplasm. Combining ability analysis of the introgressed lines further showed their potential for heterosis breeding or to be used as parental lines in breeding programme.     

Mapping polygenes for tuber resistance to late blight in a diploid Solanum phureja × S. stenotomum hybrid population

Potato tuber blight is a disease caused by the oomycete Phytophthora infestans (Mont.) de Bary. Due to the significant economic impact of this disease, introgression of durable resistance into the cultivated potato is one of the top priorities of breeding programmes worldwide. Though numerous resistance loci against this devastating disease have already been mapped, most of the detected loci are contributing towards foliar resistance while specific information on tuber resistance is limited. To identify the genetic components of tuber resistance and its relationship to foliar resistance and plant maturity we have investigated the host‐pathogen interaction in a segregating diploid hybrid Solanum phureja × S. stenotomum family. Mature tubers from this mapping family were inoculated with a sporangial suspension of P. infestans (US‐8 clonal lineage) and evaluated for lesion expansion. No significant correlation was detected between late blight resistance in foliage and tubers, and between plant maturity and tuber resistance. Four chromosomal regions were significantly associated with tuber resistance to the disease. The largest effect was detected near the marker locus PSC (LOD 10.7) located on chromosome 10. This locus explained about 63% of the total phenotypic variation of the trait. The other three resistance‐related loci were mapped on chromosomes 8 (GP1282, LOD 4.4), 6 (CP18, LOD 4.0) and 2 (CP157, LOD 3.8). None of the four tuber resistance loci coincides with the foliage resistance loci detected in this same family. Tuber blight resistance quantitative trait loci (QTL) on chromosomes 2, 8 and 10 are distinct from the maturity QTLs and have an additive effect on tuber resistance. These results indicate that different genes are involved in foliar and tuber resistance to P. infestans in the present family and that some of the resistance genes might be associated with late maturity.     

Pollen viability and meiotic behaviour in intraspecific hybrids of Hydrangea aspera subsp. aspera Kawakami group x subsp. sargentiana

Pollen viability and male meiosis in intraspecific hybrids of Hydrangea aspera subsp. aspera Kawakami group (2n = 2x = 36) and subsp. sargentiana (2n = 2x = 34) were investigated. Although it had been assumed that they were sterile, pollen viability was observed; it varied from 2.5% to 12.1%. The production of gametes with different chromosome numbers was implied by the analysis of the dispersion of the diameter distribution of pollen grains. Analysis of male meiosis made it possible to identify the origins with two major categories of meiotic aberrations: abnormal chromosome distribution (early chromosome migration at metaphase, lagging chromosomes at anaphase, micronuclei at telophase), leading to the formation of unbalanced tetrads and/or ones with supernumerary microspores; and abnormal spindle orientation in metaphase II (tripolar, fused and parallel spindles), leading to the formation of dyads or triads. The mode of 2n pollen formation is of the First Division Restitution type. The high level of parental heterozygosity that is normally associated with them should facilitate the transfer of a polygenic trait in breeding programme.     

Optimizing embryo age and media for enhancing hybrid seedling recovery in sour orange (Citrus aurantium) × Sacaton citrumelo (C. paradisi × Poncirus trifoliata) crosses through embryo rescue

In citrus, development of new hybrids is difficult due to failure of the germination of zygotic embryos. Hence, this study was conducted to standardize embryo age and media for maximizing the germination and subsequent seedling growth in sour orange × Sacaton citrumelo crosses followed by the identification of simple sequence repeats (SSR) markers for distinguishing the hybrids. A factorial experiment in complete randomized design was conducted with two each embryo age (110–120 and 130–140 days after pollination) and culture media (G‐B5 supplemented with 1.5 mg/L GA₃ plus malt extract and G‐B5 plus malt extract) for in vitro embryo culture of sour orange × Sacaton citrumelo progenies. Older embryos germinated well in both media, though highest on M1 medium (100%). In fact, M1 medium gave higher plantlet survival in both age groups. The seedling growth of older embryos had highest root length (37.80 mm) and shoot length (24.20 mm) in M1 medium at 60 days after inoculation (DAI). Three SSR markers (TAA45, CAC15 and CAC39) showed polymorphism between female and male parents and were able to identify their hybrids.     

Inheritance of aroma compounds in a model population of Fragaria × ananassa Duch.

The inheritance of important aroma compounds is demonstrated by the use of a model population of Fragaria  ×  ananassa . Two cultivars were chosen for the crossing, 'Mieze Schindler' and 'Elsanta', which differ strongly in pedigree, phenotype, and fruit traits, including flavour. Two hundred seedlings of this cross-combination were randomly selected from a population of 438 seedlings and propagated, with three plants per seedling. Aroma analysis was performed by automated headspace solid phase micro-extraction in combination with non-targeted data analysis (pattern recognition). This method can assist breeding programmes by the preparation and analysis of hundreds of samples by a simple procedure. The method was backed up by the substance identification using gas chromatography/mass spectrometry. Data processing of 78 detectable volatile compounds by Chromstat^TM in the F₁ population showed very high variability of the volatile patterns. The ester methyl anthranilate (MA) is a discriminative key compound, being present only in the mother cultivar 'Mieze Schindler', and not detectable in 'Elsanta'. Methyl anthranilate was detectable in one fourth of the F₁ population. This low degree of inheritance shows that this important compound is easily lost in the breeding process, which could explain why all investigated modern cultivars do not contain MA. The results are valuable for the choice of breeding partners and for selection in seedling populations.     

QTL analysis of quality traits in the spring wheat cross RL4452 × 'AC Domain'

Wheat grain quality is a complex group of traits of tremendous importance to wheat producers, end‐users and breeders. Quantitative trait locus (QTL) analysis studied the genetics of milling, mixograph, farinograph, baking, starch and noodle colour traits in the spring wheat population RL4452/‘AC Domain’. Forty‐seven traits were measured on the population and 99 QTLs were detected over 18 chromosomes for 41 quality traits. Forty‐four of these QTLs mapped to three major QTL clusters on chromosomes 1B, 4D, and 7D. Fourteen QTLs mapped near Glu‐B1, 20 QTLs mapped near a major plant height QTL on chromosome 4D, and 10 QTLs mapped near a major time to maturity QTL on chromosome 7D. Large QTLs were detected for grain and flour protein content, farinograph absorption, mixograph parameters, and dietary fibre on chromosome 2BS. QTLs for yellow alkaline noodle colour parameter L* mapped to chromosomes 5B and 5D, while the largest QTL for the b* parameter mapped to 7AL.     

AFLP–SSR maps of maize × teosinte and maize × maize: comparison of map length and segregation distortion

Two genetic linkage maps of Zea mays were constructed: one population comprised 94 F₂ individuals of a dent ‘B64’ × teosinte (Z. mays ssp. huehuetenangensis) cross while the second consisted of 94 F₂ individuals of a ‘B64’ × Caribbean flint ‘Na4’ cross. The level of polymorphism was higher in the ‘B64’ × teosinte combination than the ‘B64’ × ‘Na4’ combination. In the ‘B64’ × teosinte cross, a total of 338 amplified fragment length polymorphism (AFLP) and 75 simple sequence repeat (SSR) markers were mapped to 10 chromosomes, which covered 1402.4 cM. In the ‘B64’ × ‘Na4’ cross, a total of 340 AFLP and 97 SSR markers were mapped to 10 chromosomes, covering 1662.8 cM. Segregation distortion regions were found on chromosomes 4, 5 and 8 in the ‘B64’ × teosinte cross and on chromosome 9 in the ‘B64’ × ‘Na4’ cross. Comparison of the two maps revealed that the maize × teosinte map was 11.5% shorter than the maize × maize map. The maps generated in this study may be useful to identify genes controlling flooding tolerance.     

Meiosis and fertility in interspecific hybrids of Nicotiana tabacum L. ×N. glauca Grah. and their derivatives

Nicotiana glauca is of potential interest to breeders as it carries resistance to black root rot of tobacco. Cytological investigations of sexual interspecific hybrids of N. tabacum T′T′TT (2n = 4x = 48) cv. ‘Wiślica’ × N. glauca GG (2n = 2x = 24) were carried out. The analyses of chromosome association at diakinesis and metaphase I in the PMCs of amphihaploid F₁ T′TG (2n = 3x = 36) revealed low variable pairing with 0–5 bivalents. The sterile amphihaploids F₁ were converted into partial female fertile amphidiploids T′T′TTGG by chromosome doubling. Among 36 mature plants obtained, 15 were found to have chromosome numbers (2n = 6x = 72) and were verified as amphidiploids, 9 had (2n = 6x = 70 or 71) chromosomes while the remaining 12 were haploid. True amphidiploids, in spite of quite high chromosome pairing during meiosis, were very different in pollen fertility, ranging from 0% to 85%. Male fertility disturbances did not correlate with the degree of female fertility upon pollination with N. tabacum. Sesquidiploids T′TG (2n = 5x = 60) obtained from backcrossing the amphidiploids to parental tobacco showed more than 22 bivalents, 10–12 univalents and occasional multivalents that indicated the possibility of interchange events between N. tabacum and N. glauca genomes.     

Molecular mapping of soybean seed tocopherols in the cross ‘OAC Bayfield’ × ‘OAC Shire’

Soybean (Glycine max [L.] Merr.) is cultivated primarily for its protein and oil in the seed. In addition, soybean seeds contain nutraceutical compounds such as tocopherols (vitamin E), which are powerful antioxidants with health benefits. The objective of this study was to identify molecular markers linked to quantitative trait loci (QTL) that affect accumulation of soybean seed tocopherols. A recombinant inbred line (RIL) population derived from the cross ‘OAC Bayfield’ × ‘OAC Shire’ was grown in three locations over 2 years. A total of 151 SSR markers were polymorphic of which a one‐way analysis of variance identified 42 markers whereas composite interval mapping identified 26 markers linked to tocopherol QTL across 17 chromosomes. Individual QTL explained from 7% to 42% of the total phenotypic variation. Significant two‐locus epistatic interactions were identified for a total of 122 combinations in 2009 and 152 in 2010. The multiple‐locus models explained 18.4–72.2% of the total phenotypic variation. The reported QTL may be used in marker‐assisted selection (MAS) to develop high tocopherol soybean cultivars.     

Cytological behaviour of the somatic hybrids Passiflora edulis f. flavicarpa + P. cincinnata

Meiosis and pollen viability of interspecific somatic hybrids, i.e. Passiflora edulis f. flavicarpa , the yellow passion fruit +  P. cincinnata , a wild species, were examined and compared with their corresponding diploid fusion parents. Firstly, the cybrid nature of these hybrids was confirmed by leaf-stomatal measurements and RFLP analyses using four mitochondrial genes as probes. The meiotic behaviour revealed relatively high stability, with most of the hybrid cells showing 18 bivalents. Some instability, such as a quadrivalent configuration was also recorded which has been interpreted as an interchange that occurred in the progenitors more than as a result of in vitro culture or chromosome reorganization in the new genome. Even in low frequencies, the occurrence of univalents resulted in mis-division, laggard and micronucleus formation. High values of pollen viability (>70%) were found in the diploid parents as well as in the hybrid plants. The results are discussed in relation to the possibilities of applying somatic hybridization for improving passion fruit varieties.     

Pod shatter resistance evaluation in cultivars and breeding lines of Brassica napus, B. juncea and Sinapis alba

Pod shatter susceptibility was investigated in Brassica napus germplasm and shatter resistant species of B. juncea and Sinapis alba. The comparisons were made by measuring seed yield in field plots, detached pod rupture energy (RE) and the half‐life of pod‐opening. Pod shatter resistance was significantly greater in B. napus lines derived from interspecific hybridizations of B. napus with B. rapa, B. carinata and B. juncea, than common B. napus cultivars. While these lines exhibited no significant difference in resistance to pod shatter than B. juncea, an entry of S. alba had no yield loss caused by pod shatter. Resistance to pod shatter was characterized in the field as little or no yield loss after full maturity, delayed shattering in time, and stable yield performance under variable climatic conditions during pod maturity. Yield loss caused by pod shatter ranged from a low of 4% for the B. juncea cv. ‘AC Vulcan’ to a high of 61% for the black seeded B. napus line DH12075 in 2‐year field trials after 1 month maturity. Pod shatter resistance was not significantly associated with specific plant and pod morphological traits, except pod length (P = 0.005) in tested materials. Field visual scores of pod shatter through inspections of average pod shatter per plant within plots were highly correlated with plot yield loss. Indoor quantitative evaluations of pod strength using a pendulum machine to measure pod RE and random impact test to measure half‐life of pod‐opening resistance were highly correlated with field yield loss. Multiple evaluations of pod shatter in method and in time after pod maturity are recommended for reliable evaluation of pod shatter resistance.     

The host background of rice influences the resistance expression of a three genes pyramid (xa5 + xa13 + Xa21) to bacterial blight (Xanthomonas oryzae pv. oryzae) pathotypes of Indian mainland and Bay islands

Bacterial blight (BB) is a major disease of rice for which host resistance is the only effective solution. The three genes pyramid xa5 + xa13 + Xa21 is recently the most utilized combination for developing resistant varieties through marker‐assisted breeding. Our study was carried out to elicit the detailed response of twenty lines possessing these three genes in five genetic backgrounds to twelve diverse BB pathotypes in India. The lines developed from ADT 47 variety showed incomplete resistance to most of the pathotypes, whereas susceptibility varied from 8.3% to 16.6% in ADT 43 and IR24, respectively. However, in IMP ASD16/60 and Improved Samba Mahsuri, complete resistance against all pathotypes was observed. The overall results confirmed that genetic background plays crucial role for the effective expression of xa5 + xa13 + Xa21 combination. Molecular studies did not reveal correlation between origin of pathotypes and their virulence potential. It is suggested to deploy Improved Samba Mahsuri, IMP ASD 16/60 and AD1306 varieties in the bacterial blight prone areas or use them as donors for realizing wider and durable resistance.     

Quantitative trait loci mapping of adult‐plant resistance to leaf rust in a Fundulea 900 × ‘Thatcher’ wheat cross

Wheat leaf rust (LR), caused by the obligate biotrophic fungus Puccinia triticina (Pt), is a destructive foliar disease of common wheat (Triticum aestivum L.) worldwide. The most effective, economic means to control the disease is resistant cultivars. The Romanian wheat line Fundulea 900 showed high resistance to LR in the field. To identify the basis of resistance to LR in Fundulea 900, a population of 188 F_2:3 lines from the cross Fundulea 900/‘Thatcher’ was phenotyped for LR severity during the 2010–2011, 2011–2012 and 2012–2013 cropping seasons in the field at Baoding, Hebei Province. Bulked segregant analysis and simple sequence repeat markers were used to identify the quantitative trait loci (QTLs) for LR adult‐plant resistance in the population. Three QTLs were detected and designated as QLr.hebau‐1BL, QLr.hebau‐2DS and QLr.hebau‐7DS. Based on the chromosome positions and molecular marker tests, QLr.hebau‐1BL is Lr46, and QLr.hebau‐7DS is Lr34. QLr.hebau‐2DS was derived from ‘Thatcher’ and was close to Lr22. This result suggests that Lr22b may confer residual resistance on field nurseries when challenged with isolates virulent on Lr22b, or another gene linked to Lr22b confers this resistance from ‘Thatcher’. This study confirms the value of Lr34 and Lr46 in breeding for LR resistance in China; the contribution of the QTL to chromosome 2D needs further validation.     

QTL mapping of adult‐plant resistance to stripe rust in a ‘Lumai 21 × Jingshuang 16’ wheat population

Stripe rust, caused by Puccinia striiformis f. sp. tritici, is a devastating fungal disease in common wheat (Triticum aestivum L.) worldwide. Chinese wheat cultivars ‘Lumai 21’ and ‘Jingshuang 16’ show moderate levels of adult‐plant resistance (APR) to stripe rust in the field, and they showed a mean maximum disease severity (MDS) ranging from 24 to 56.7% and 26 to 59%, respectively, across different environments. The aim of this study was to identify quantitative trait loci (QTL) for resistance to stripe rust in an F₃ population of 199 lines derived from ‘Lumai 21’ × ‘Jingshuang 16’. The F₃ lines were evaluated for MDS in Qingshui, Gansu province, and Chengdu, Sichuan province, in the 2009–2010 and 2010–2011 cropping seasons. Five QTL for APR were detected on chromosomes 2B (2 QTL), 2DS, 4DL and 5DS based on mean MDS in each environment and averaged values from all three environments. These QTL were designated QYr.caas‐2BS.2, QYr.caas‐2BL.2, QYr.caas‐2DS.2, QYr.caas‐4DL.2 and QYr.caas‐5DS, respectively. QYr.caas‐2DS.2 and QYr.caas‐5DS were detected in all three environments, explaining 2.3–18.2% and 5.1–18.0% of the phenotypic variance, respectively. In addition, QYr.caas‐2BS.2 and QYr.caas‐2BL.2 colocated with QTL for powdery mildew resistance reported in a previous study. These APR genes and their linked molecular markers are potentially useful for improving stripe rust and powdery mildew resistances in wheat breeding.