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.     

Pollen viability and stigma receptivity in <Emphasis Type="Italic">Lilium</Emphasis> during anthesis

Pollen viability and stigma receptivity are prerequisites for successful pollination and seed set in flowering plants. In this study, the pollen viabilities and stigma receptivities of nine Lilium genotypes (six cultivars and three species native to China) were assayed by in vitro pollen germination and the benzidine-H₂O₂ method, respectively. Embryo sac development during anthesis was observed to further ensure the timing of controlled pollination. In addition, the relationship between stigma secretion and stigma receptivity was studied to estimate the pollination time based on phenotype. Anthers cracked on the day of flowering in all genotypes, but pollen germination during anthesis was not observed in Asiatic hybrids excepted for ‘Tiny pudhye’, which exhibited low pollen viability for a short period of time (from 0 to 1 day after anthesis). In the other genotypes, pollen germination rates were highest on anthesis (five of seven genotypes), 0–1 day after anthesis (L. sulphureum), or 0–2 days after anthesis (one Longiflorum hybrid), and then gradually decreased with days after anthesis. While, stigma receptivity first increased and then decreased during anthesis. For most genotypes, stigmas began to be receptive 1 day after anthesis, and all genotypes exhibited stigma receptivity at 2 days after anthesis. The durations of stigma receptivity and strongest stigma receptivity, were genotype dependent, and were 5–8 days and 1–4 days, respectively. Moreover, on the first flowering day, 6 of 7 genotypes had mature embryo sacs, and at the time at which stigmas began to be receptive, all tested genotypes had mature embryo sacs. Some Lilium genotypes showed stigma secretion, which can be a sign of stigma receptivity. Stigmas became receptive and reached highest receptivity within 1 day of the first appearance of secretion on the surface of the stigma and at peaking, respectively. The results of this study are valuable for the implementation of successful Lilium breeding programs.     

Pollination and in vitro germination of seeds for interspecific hybridization of <Emphasis Type="Italic">Psidium guajava</Emphasis> and <Emphasis Type="Italic">Psidium cattleianum</Emphasis>

The genus Psidium includes important fruit crops. However, there are very few studies focusing on its reproductive biology, which limits the establishment of breeding programs. The present work investigated the reproductive biology of Psidium guajava and Psidium cattleianum in terms of compatibility of crossings between these two species aiming at interspecific hybridization because the latter species is an important source of resistance against the nematode Meloidogyne enterolobii. Several types of crosses were performed to understand the reproductive biology of these species, including the compatibility of intra- and interspecific crossings, using assisted in vivo germination of pollen grains on the stigma. In addition, the in vitro germination of both Psidium species was studied at different stages of fruit development to rescue young seeds to improve the chances of obtaining the hybrids. No fruits of 270 pollinations were obtained on guava buds at the pre-anthesis stage, regardless of the source of the pollen grain and the cultivar used as female genotype. Microscopic analyzes demonstrated the germination of pollen grains and pollen tube growth at crosses between guava cv. ‘Pedro Sato’ (P. guajava) and Psidium cattleianum. High germination percentages of Psidium cattleianum seeds were obtained in MS medium without sucrose or containing 15 g/L of this carbohydrate.     

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.     

Identifying apomixis in matroclinal progeny from an interspecific crossing between <Emphasis Type="Italic">Iris domestica</Emphasis> and three different colors of <Emphasis Type="Italic">Iris dichotoma</Emphasis>

In a previous investigation on the reciprocal difference of interspecific hybridization between three different flower colors of Iris dichotoma and Iris domestica in the F₁ offspring from crosses where I. domestica was a maternal parent were similar in morphological and cytological characters to their maternal parent. This could be evidence of apomixis; however, matroclinal progeny with complete morphological similarity to the maternal parent could be attributed to the heterozygosity for these characters in the pollen parent. The F₁ plants were investigated in order to identify apomixis in I. domestica. Four matroclinal plants were randomly selected from each F₁ population produced from Iris domestica × Iris dichotoma that had three different colors of flowers and were allowed to self-pollinate to establish an F₂ population. All of the F₂ plants had no segregation to I. domestica in their morphological characters. In addition, 13 reciprocal F₁ plants were detected by 25,719 single nucleotide polymorphism (SNP) markers. When I. dichotoma plants with three different flower colors were used as maternal parents, all the progenies were genuine hybrids. When I. domestica were used as maternal parents, all the F₁ plants were apomictic progenies. Apomixis of I. domestica was successfully identified and SNP markers identified F₁ hybrids derived from six interspecific crosses between I. dichotoma and I. domestica, which provides a reference for authenticating offspring identity during Iris cross breeding in the future.     

Slow-growing pollen-tube of colchicine-induced 2n pollen responsible for low triploid production rate in <Emphasis Type="Italic">Populus</Emphasis>

To reveal the mechanism of low triploid production, meiotic observation, induction of 2n pollen by colchicine, hybridization of colchicine-induced 2n pollen and pollen germination were conducted in Populus tomentosa clone 5088. Meiosis of the PMCs was initiated 24 h after it was cultured in the greenhouse. Results showed that the meiosis developed in a consecutive, asynchronous process. Analysis of the occurrence rate of colchicine-induced 2n pollen using the GLM-univariate revealed significant differences among the dominant meiotic stages (F = 45.822, P = 0.000). Significant differences also occurred among various colchicine injection times (F = 10.150, P = 0.004). The most effective stage for colchicine-induced 2n pollen occurred between the zygotene to diakinesis substages. Sixty-eight triploids were confirmed with both flow cytometric analysis and chromosome number-counting among the 3346 offspring. The highest triploid induction rate was 13.2%—significantly lower than the expected triploid production rate of 47.0%—indicating that the colchicine-induced 2n pollen was weak during fertilization. We found positive correlation between the efficiency of triploid production and the frequency of colchicine-induced 2n pollen (r = 0.961, P = 0.001) which suggests that the triploid production rate increased as the occurrence rate of colchicine-induced 2n pollen increased. Analysis using the GLM-univariate indicated that both pollen types (F = 87.50, P < 0.001) and durations of pollen germination (F = 7.61, P = 0.002) did have significant effects on the lengths of pollen-tubes. Haploid pollen-tubes were significantly longer than colchicine-induced 2n pollen-tubes. This suggests that haploid pollen has an advantage during fertilization.     

Genetic effect of <Emphasis Type="Italic">Striga</Emphasis> resistance in sorghum genotypes

Striga is an important parasitic weed causing substantial economic losses in cereal and legume crop production in sub-Saharan Africa. Integrated Striga management approaches such as a combined use of Striga resistant varieties and Fusarium oxysporum f.sp. strigae (FOS), a biocontrol agent of Striga, are an option to control the parasite and to boost sorghum productivity. Understanding host gene action influencing Striga resistance, with or without FOS treatment, is key to developing improved sorghum varieties with durable resistance and high yield. The objective of this study was to determine the gene action and inheritance of Striga resistance using genetically diverse populations of sorghum involving FOS treatment. Twelve sorghum parents selected for Striga resistance, FOS compatibility or superior agronomic performances were crossed using a bi-parental mating scheme. The selected male and female parents and their F₁ progenies, backcross derivatives and the F₂ segregants were field evaluated at three locations in Tanzania known for their severe Striga infestations using a lattice experimental design with two replications. The following data were collected and subjected to generation mean analysis (GMA): days-to-50% flowering (DFL), seed yield per plant (SYP) and number of Striga per plant (SN). GMA showed the preponderance of additive genetic action contributing to the total genetic variation in the evaluated sorghum populations. The additive genetic effect for DFL, SYP and SN, with and without FOS treatments, ranged from 72.02 to 86.65% and 41.49 to 95.44%, 75.62 to 91.42% and 71.83 to 91.89%, and 77.35 to 93.56% and 72.86 to 95.84%, in that order. The contribution of non-additive genetic effects was minimal and varied among generations. FOS application reduced DFL and SN and improved SYP in most of the tested sorghum populations. DFL of sorghum populations was reduced by a mean of 8 days under FOS treatment compared to the untreated control in families such as 675 × 654, AS435 × AS426 and 1563 × AS436. FOS treatment improved SYP with a mean of 6.44 g plant^?1 in 3424 × 3993 and 3984 × 672. The numbers of Striga plants were reduced with a mean of 16 plants due to FOS treatment in the crosses of 675 × 654, 1563 × AS436, 4567 × AS424, and 3984 × 672. The study demonstrated that additive genes were predominantly responsible for the inheritance of Striga resistance in sorghum. Pure line cultivar development targeting reduced DFL, SN and high SYP in the selected populations may provide enhanced response to selection for integrated Striga management (ISM) programme.     

Phenotypic variations and heritability of bearded iris breeding

Bearded iris (Iris × hybrida Hort.) refers to the large population of iris hybrids, which is identified by thick, bushy ‘beards’ on each falls (lower petals) of the blossoms. However, the selection of iris is extremely limited due to the lack of information about genotype and phenotype. According to North Carolina Design II (NC II), a statistical method to estimate the combining ability, genetic variance components and heritability, seven male parents and two female parents were hybridized to generate 14 populations of first filial generation (F₁). Each F₁ family was generated through artificial pollination in 3 years and included 60 individuals. 11 key ornamental characters, including plant height (PH), length of leaf (LL), number of flower per scape (NFS), height of individual flower (HF), diameter of flower (DF), length of fall (LF) and width of fall (WF), were measured, and some genetic parameters (heterosis, heterobeltiosis, combining ability, heritability as well as genetic correlations) were evaluated. The results showed that HF and LW had the largest number of F₁ populations gaining heterosis, thus more likely to generate taller progenies with larger blossoms. The estimates of combing ability revealed that I. ‘White and Blue’(M1) and I. ‘Sugar Blues’(M5) appeared to be ideal pollen donators. The combination of I. ‘India Chief’ × I. ‘Champagne Elegance’ (FM1 × M6) exhibited positive special combining ability (SCA) on all the tested characters, becoming the most potential populations of all. WF and NFS had the highest broad-sense heritability (91.47%) and strict-sense heritability (53.96%), respectively. Correlation analysis showed that PH was closely associated with NBS and NFS, while LL had significant correlations with HF and LF, making it possible to select taller individuals in the vegetative stage to obtain those with abundant flowers.     

Mapping of new resistance (<Emphasis Type="Italic">Vr2</Emphasis>, <Emphasis Type="Italic">Rm1</Emphasis>) and ornamental (<Emphasis Type="Italic">Di2</Emphasis>, <Emphasis Type="Italic">pl</Emphasis>) Mendelian trait loci in peach

Peach powdery mildew is one of the major diseases of the peach. Various sources of resistance to PPM have thus been identified, including the single dominant locus Vr2 carried by the peach rootstock ‘Pamirskij 5’. To map Vr2, a linkage map based on microsatellite markers was constructed from the F₂ progeny (WP²) derived from the cross ‘Weeping Flower Peach’ × ‘Pamirskij 5’. Self-pollinations of the parents were also performed. Under greenhouse conditions, all progenies were scored after artificial inoculations in two classes of reactions to PPM (resistant/susceptible). In addition to Vr2, WP² segregated for three other traits from ‘Weeping Flower Peach’: Rm1 for green peach aphid resistance, Di2 for double-flower and pl for weeping-growth habit. With their genomic locations unknown or underdocumented, all were phenotyped as Mendelian characters and mapped: Vr2 mapped at the top of LG8, at 3.3 cM, close to the CPSCT018 marker; Rm1 mapped at the bottom of LG1, at a position of 116.5 cM, cosegregating with the UDAp-467 marker and in the same region as Rm2 from ‘Rubira’^®; Di2 mapped at 28.8 cM on LG6, close to the MA027a marker; and pl mapped at 44.1 cM on LG3 between the MA039a and SSRLG3_16m46 markers. Furthermore, this study revealed, for the first time, a pseudo-linkage between two traits of the peach: Vr2 and the Gr locus, which controls the red/green color of foliage. The present work therefore constitutes a significant preliminary step for implementing marker-assisted selection for the four major traits targeted in this study.     

Morphological and molecular characterization of the second backcross progenies of Ogu-CMS Chinese kale and rapeseed

Ogura cytoplasmic male sterility (Ogu-CMS) is widely used in the production of commercial hybrids of Brassica oleracea. However, the widespread application of the Ogu-CMS system in B. oleracea has hindered the germplasm innovation of Ogu-CMS resources due to the lack of a natural restorer line. Previously, the Ogu-CMS fertility-restored interspecific hybrids between rapeseed 15Y403 (2n = 38, AACC) and Chinese kale JL1 (2n = 18, CC) have been successfully produced. However, these progenies, which still contained a large proportion of rapeseed genomic components, showed poor fertility and a low seed setting rate under natural pollination. To improve fertility and seed setting, a successive backcross with JL1 was performed to produce BC₂ progenies. Screening with the Rfo-specific marker, five individuals harboring the Rfo gene were identified among 98 BC₂ progenies. These five individuals underwent background marker screening and an evaluation of agronomic traits and fertility. One individual (code: 15Q23) was identified with higher pollen viability, better seed setting under natural pollination, and a closer genetic background to the parent Chinese kale JL1. Many morphological traits showed no significant differences (P < 0.05) between 15Q23 and the backcross parent JL1. However, the average seed setting of 15Q23 under natural pollination was 0.72 seeds per pod, which was 50 times higher than that of BC₁ progenies, and the average pollen viability was 87.07%, which was significantly better than that of the F₁ and BC₁ plants (P < 0.01). The genetic background of 15Q23 was more closer to the parent JL1 than that of BC₁ plants and another BC₂ fertility-restored individual, with 82% of the polymorphic alleles being the same as those of the parent Chinese kale JL1. Thus, the individual 15Q23 could be used as a donor plant for further backcrosses. This study lays the foundation for the development of Ogu-CMS restorer material in B. oleracea.     

Transformation of <Emphasis Type="Italic">Campanula</Emphasis> by wild type <Emphasis Type="Italic">Agrobacterium rhizogenes</Emphasis>

Compact growth is an important quality criterion in horticulture. Many Campanula species and cultivars exhibit elongated growth which is suppressed by chemical retardation and cultural practice during production to accommodate to the consumer’s desire. The production of compact plants via transformation with wild type Agrobacterium rhizogenes is an approach with great potential to produce plants that are non-GMO. Efficient transformation and regeneration procedures vary widely among both plant genera and species. Here we present a transformation protocol for Campanula. Hairy roots were produced on 26–90% of the petioles that were used for transformation of C. portenschlagiana (Cp), a C. takesimana × C. punctata hybrid (Chybr) and C. glomerata (Cg). Isolated hairy roots grew autonomously and vigorously without added hormones. The Cg hairy roots produced chlorophyll and generated plantlets in response to treatments with cytokinin (42 µM 2iP) and auxin (0.67 µM NAA). In contrast, regeneration attempts of transformed Cp and Chybr roots lead neither to the production of chlorophyll nor to the regeneration of shoots. Agropine A. rhizogenes strains integrate split T-DNA in T_L- and T_R-DNA fragments into the plant genome. In this study, regenerated plants of Cg did not contain T_R-DNA, indicating that a selective pressure against this T-DNA fragment may exist in Campanula.     

Development of late-bolting F<Subscript>1</Subscript> hybrids of Chinese cabbage (<Emphasis Type="Italic">Brassica rapa</Emphasis> L.) allowing early spring cultivation without heating

We developed new F₁ hybrids of Chinese cabbage (Brassica rapa L. ssp. pekinensis) that allow cultivation earlier in spring without heating by introducing extremely late-bolting alleles at two homologs of the flowering repressor Flowering Locus C (BrFLC2 and BrFLC3) from non-heading ‘Leafy Green Parental Line No. 2’. These new F₁ hybrids were produced by the following four steps. First, the extremely late bolting selected lines were developed. These selected lines headed in spring after overwintering cultivation, whereas the conventional F₁ cultivars flowered. Secondly, an investigation of the three plantings showed that our F₁ hybrids formed heads when seeds were sown from mid-February to early March, whereas the conventional F₁ cultivar did not form heads because of premature bolting. Thirdly, we identified some F₁ hybrids with extremely late bolting during early spring cultivation in an investigation of many F₁ hybrids. Finally, based on an investigation across four cold regions for 2 years, we compared the commercialization rate, defined as the proportion of plants greater than 2000 g in weight and with a flowering stalk less than 10 cm long. Then we identified a F₁ of MS02 × 12-04 which had a high commercialization rate on average (92%), whereas the rates of three conventional F₁ cultivars were only 0–2%. In the near future, these F₁ hybrids will be valuable late-bolting cultivars despite climate change, permitting stable cultivation and harvest over wide regions.     

Genetic mapping of resistance to <Emphasis Type="Italic">Puccinia hordei</Emphasis> in three barley doubled-haploid populations

The success of breeding for barley leaf rust (BLR) resistance relies on regular discovery, characterization and mapping of new resistance sources. Greenhouse and field studies revealed that the barley cultivars Baronesse, Patty and RAH1995 carry good levels of adult plant resistance (APR) to BLR. Doubled haploid populations [(Baronesse/Stirling (B/S), Patty/Tallon (P/T) and RAH1995/Baudin (R/B)] were investigated in this study to understand inheritance and map resistance to BLR. The seedlings of two populations (B/S and R/B) segregated for leaf rust response that conformed to a single gene ratio (\({\text{X}}_{1:1}^{2}\) = 0.12, P > 0.7 for B/S and \({\text{X}}_{1:1}^{2}\) = 0.34, P > 0.5 for R/B) whereas seedlings of third population (P/T) segregated for two-gene ratio (\({\text{X}}_{1:1}^{2}\) = 0.17, P > 0.6) when tested in greenhouse. It was concluded that the single gene in Baudin and one of the two genes in Tallon is likely Rph12, whereas gene responsible for seedling resistance in Stirling is Rph9.am (allele of Rph12). The second seedling gene in Tallon is uncharacterized. In the field, APR was noted in lines that were susceptible as seedlings. A range of disease responses (CI 5–90) was observed in all three populations. Marker trait association analysis detected three QTLs each in populations B/S (QRph.sun-2H.1, QRph.sun-5H.1 and QRph.sun-6H.1) and R/B (QRph.sun-1H, QRph.sun-2H.2, QRph.sun-3H and QRph.sun-6H.2), and four QTLs in population P/T (QRph.sun-6H.2, QRph.sun-1H.2, QRph.sun-5H.2 and QRph.sun-7H) that significantly contributed to low leaf rust disease coefficients. High frequency of QRph. sun-5H.1, QRph. sun-6H.1, QRph. sun-1H.1, QRph. sun-2H.2, QRph. sun-6H.2, QRph. sun-7H (based on presence of the marker, closely associated to the respective QTLs) was observed in international commercial barley germplasm and hence providing an opportunity for rapid integration into breeding programmes. The identified candidate markers closely linked to these QTLs will assist in selecting and assembling new APR gene combinations; expectantly this will help in achieving good levels of durable resistance for controlling BLR.     

The sporophytic self-incompatibility mating system is conserved in <Emphasis Type="Italic">Olea europaea</Emphasis> subsp. <Emphasis Type="Italic">cuspidata and O. e. europaea</Emphasis>

Fruit setting after self-pollination, crosses and free-pollination appears to be erratic in the cultivated olive tree [Olea europaea subsp europaea L. (O. e. europaea L.)] because of a lack of a suitable model to enable prediction of rates. The same lack of prediction also applies to the wild taxon Olea europaea subsp cuspidata (O. e. cuspidata). Because of their close phylogenetic relationships, we hypothesize that O. e. cuspidata and cultivated olive share the same self-incompatibility system. We used data recently published in a wide study involving four O. e. cuspidata accessions and four olive cultivars. Because the olive varieties have been deciphered for their S-allele pair, that infer determinants present in the stigma and pistil, and that coat the pollen, we deciphered the S-alleles carried by three of the O. e. cuspidata accessions. Data are too scarce and the number of accessions too small to speculate on the O. e. cuspidata genetic population structure. The working hypothesis is confirmed. This study and data from the Italian team will enable us to embark on a large-scale hybridization program between the two subsp. to obtain a wide range of progenies for screening for responses to cold, diseases and pests.     

Karyotype analysis of eight wild <Emphasis Type="Italic">Tulipa</Emphasis> species native to China and the interspecific hybridization with tulip cultivars

The information of ploidy, karyotype and genetic relationship is useful for interspecific hybridization in ornamental plants. For Tulipa species native to China, very limited cytological information is available now. The objective of this study was to verify the chromosome number, karyotype and genetic relationship of the eight Tulipa species: T. edulis, T. schrenkii, T. iliensis, T. thianschanica, T. altaica, T. sinkiangensis, T. heterophylla and T. buhseana. And the interspecific crosses were made between T. altaica and ten tulip cultivars to obtain novel germplasm. The ovary-swelling, fruit-setting and bulblet formation rates were surveyed when different ploidy cultivars were used as female parents. This work confirmed that all eight species collected in China were diploid (2n?=?2x?=?24), among which chromosome numbers of T. thianschanica, T. sinkiangensis and T. heterophylla were firstly reported and the karyotypes of all any other species except for T. edulis were determined for the first time. The karyotypes of eight Tulipa species were classified as 3A, 4A or 3B. The results of interspecific hybridization showed significant difference when different ploidy cultivars were used as female parents. The highest fruit-setting rate was obtained when diploid cultivars were used as female parents crossed with diploid T. altaica, whereas the ovary swelling was observed in two out of four triploid cultivars as female parents, and no seeds were harvested when tetraploid cultivars were used as female parents. Our findings provided an effective means of cultivar improvement in tulip.     

Characterization of the resistance to <Emphasis Type="Italic">Phytophthora infestans</Emphasis> in local potato cultivars in Bolivia

This experiment was carried out to investigate whether and how much field resistance to late blight, caused by Phytophthora infestans, is present in the local cultivated potato germplasm. In total 36 entries were compared in a field experiment in an area highly conducive to late blight development. Of the 36 cultivars 32 were local cultivars belonging to five Solanum species, S. tuberosum (1 accession), S. andigena (18), S. juzepczukii (2), S. stenotomum (9) and S. ajanhuiri (2). The other four cultivars were derived from breeding programmes, one being the Dutch cultivar Alpha used as a highly susceptible control. The 36 cultivars were planted according to a simple 6 × 6 lattice design with three replicates. Each replicate was divided in six incomplete blocks each with six cultivars. The disease severity was assessed weekly during 9 weeks starting 48 days after planting. The area under the disease progress curve (AUDPC) was used as a measure of the field resistance. Nine isolates from surrounding potato fields were tested for their virulence to the resistance genes R1–R11 using 22 differential cultivars. The components of the field resistance of 19 of these cultivars were compared in the greenhouse using a local isolate with virulence to all known R-genes, except to R9. The nine isolates represented seven races with a race complexity varying from 7 to 10 virulence factors. All isolates carried virulence against R1, R2, R3, R7, R10 and R11, while virulence against R9 was absent. The AUDPC among the 32 local cultivars ranged from very large, significantly larger than that of ‘Alpha’ to very small. The AUDPC from S. stenotomum accessions ranged from very large to intermediate, those from S. andigena accessions from large to very small. Especially among the S. andigena accessions interesting levels of field resistance were found. Four components of field resistance were assessed, latency period (LP), lesion size (LS), lesion growth rate (LGR) and relative sporulation area (RSA). All four showed a considerable variation among the cultivars. The LP ranged from 3½ to 6 days. The LS ranged from 225 mm² to 20 mm². The LGR varied about six-fold, the RSA more than 10-fold. The components tended to vary in association with one another. LP and LGR were well associated with each other and had a significant correlation with the AUDPC.     

<Emphasis Type="Italic">In vitro</Emphasis> plant regeneration,flowering and fruiting from nodal explants of <Emphasis Type="Italic">Andrographis lineata</Emphasis> nees (Acanthaceae)

In the present study, in vitro propagation of tribal endemic medicinal plant, Andrographis lineata has been established using mature nodal explants. High frequency of regeneration (91.4%) was achieved on MS medium containing BA (3.0 mg L^–1) along with IAA (0.2 mg L^–1). Strikingly, irrespective of the season and collection period, we observed axillary flower induction and fruit formation from the above in vitro cultures supplemented with BA and NAA. Transition from the vegetative to the reproductive phase occurred within 2 months of culture, and importantly was influenced by factors such as sucrose and cytokinins. In vitro-regenerated flowers were morphologically identical to in vivo flowers. Furthermore, our scanning electron microscopic studies revealed that pollen external morphology of both in vitro and in vivo flowers were similar. The flowers self-fertilized and produced fruits in vitro. Elongated shoots rooted well on half-strength MS basal medium, and were successfully acclimatized to the garden conditions. Altogether, our established protocol can be utilized in plant breeding for the purpose of ex situ conservation, quick flowering, and fruit set.     

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.     

QTL Identification and Fine Mapping for Seed Storability in Rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.)

Rice seed storability is an important characteristic of seed quality so that the cultivars with strong seed storability are expected in the production of hybrid seeds. Presently, little is known about the genetic and physiological mechanisms controlling rice seed storability. In this study, a double haploid population derived from the cross between a japonica cultivar CJ06 and an indica cultivar TN1 was used to identify the quantitative trait loci (QTLs) for seed germination percentage (GP) and fatty acid content (FA) during natural storage or artificial aging. A total of 19 QTLs, including ten QTLs for GP and nine QTLs for FA, were identified on nine chromosomes with the phenotypic variations ranged from 2.1 to 22.7%. Besides, six and three pairs of epistatic interactions were identified for GP and FA, respectively. Moreover, qGP-9, a QTL for germination percentage, was delimited in an interval of 92.8 kb between two STS markers P6 and P8, which contains 15 putative open reading frames. These results provide important information for understanding the genetic mechanisms on rice seed storability, and will be useful for breeding new rice varieties with high seed storability.     

Relationship between hybrid performance and genetic variation in self-fertile and self-sterile sugar beet pollinators as estimated by SSR markers

Sugar beet hybrid varieties are produced through the crosses between male sterile lines and the multigerm pollinators. The uniformity of pollinators used for hybrid crosses depends on the presence of self-sterility (S ^s ) and self-fertility (S ^f ) genes. The aim of the study was to analyze correlation between hybrid performance and genetic distance or heterozygosity of the sugar beet pollinators. Twelve diploid pollinators classified as self-sterile (S ^s ) or self-fertile (S ^f ) and two cytoplasmic male sterile (CMS) lines were crossed in line × tester scheme, producing 24 F₁ hybrids. The parents and the hybrids were evaluated for root yield and quality traits, from which F₁ performance, combining abilities, mid-parent and high-parent heterosis were calculated. Parental genetic distance and diversity of the pollinators were estimated by SSR markers and, together with GCA and F₁ performance, correlated with the heterosis effects. The S ^f hybrids had better GCA and higher values of root yield, root weight, and root circumference than the S ^s hybrids. Heterosis was recorded in more combinations with the S ^f than with the S ^s pollinators. Parameters of genetic diversity were higher in the S ^s (Na = 3.125; Ne = 2.341; He = 0.555) than in the S ^f pollinators (Na = 3.000; Ne = 2.188; He = 0.510). Genetic distance between the tested pollinators and the CMS lines was low (0.072–0.224) indicating that the genetic base of the investigated germplasm was narrow. Correlation of the heterosis effects with GD and heterozygosity was detected only for the root yield traits.