F1 Hybrid Progeny Research Articles

#966 Targeting cAMP in ADPKD by small molecule activation of long form PDE4 enzymes

Abstract Background and Aims Autosomal Dominant Polycystic Kidney Disease (ADPKD) is the most common hereditary kidney disorder, affecting over 12 million people worldwide. ADPKD is characterized by the uncontrolled growth of fluid-filled cysts in the kidney that can lead to kidney enlargement, loss of function, and associated complications. Currently, there are limited treatment options for ADPKD patients. By activating long form PDE4 enzymes and increasing cAMP hydrolysis, Mironid®’s first-in-class small molecule LoAc® compounds [1] directly target the increased kidney cAMP levels that are known to drive cyst formation in ADPKD, resulting in decreased cyst growth in vitro and in vivo. Here we present translational data demonstrating that LoAc® compound MR-L22 lowers renal cAMP after single or multiple doses and delivers efficacy in the Pkd1RC/RC mouse model of ADPKD, alongside favourable differentiation from tolvaptan, the only therapy approved for ADPKD patients. Method To demonstrate cAMP lowering in urine, male Han Wistar rats (n = 6) were dosed p.o. with 100 mg/kg MR-L22 or vehicle (1% methylcellulose, 0.2% Tween 80) either once, or on 7 consecutive days. Urine was collected on Day 1 or Day 7. To assess the ability of MR-L22 to ameliorate disease progression alone, or in combination with tolvaptan, doses of MR-L22 (60 mg/kg) and tolvaptan (0.1% in chow), both expected to be individually sub-maximally efficacious, were administered singly, or in combination in F1 hybrid progeny Pkd1RC/RC mice (8 m and 8f/group), from 4-16 weeks of age. Urine measurements were taken after 8 weeks of dosing. Total kidney volume (TKV) was measured by MRI prior to randomisation and termination. cAMP in urine and kidney tissue were measured by ELISA. Results A 100 mg/kg oral dose of MR-L22 in rat resulted in a >40% decrease in cAMP in urine compared to vehicle. This effect was maintained over multiple days of dosing demonstrating that urinary cAMP can be used as a biomarker of LoAc® target engagement. Consistent with the mechanism of action, a small increase in urine production and commensurate decrease in urine osmolality was observed. To assess the ability of MR-L22 to ameliorate disease progression in an in vivo model of ADPKD, and to determine whether additive or synergistic effects might be seen in combination with tolvaptan, MR-L22 (60 mg/kg) and tolvaptan (0.1% in chow) were dosed individually and in combination in the Pkd1RC/RC mouse model of ADPKD. Alongside a reduction in urinary cAMP, MR-L22 delivered reductions in TKV/Bw (Fig. 1), urinary and tissue cAMP, cystic index, terminal blood urea nitrogen (BUN) and creatinine, as well as trending reductions in urinary NGAL. Conclusion We have shown that in rat, acute or multiple dosing, of LoAc® compound MR-L22 results in a significant reduction of urinary cAMP and a small, but measurable, increase in urine production and commensurate decrease in urine osmolality. Furthermore, we have demonstrated that significantly reduced urinary and kidney cAMP levels correlate with reductions in TKV, cystic index, Kw/Bw, BUN and urinary NGAL in the Pkd1RC/RC model of ADPKD. In these studies, we have demonstrated beneficial differentiation vs tolvaptan on limiting aquaretic effects but been unable to conclusively demonstrate additive or synergistic effects of MR-L22 combined with tolvaptan due to experimental limitations; Further experiments will be needed to answer this question. Looking forwards, measurements of urine cAMP and urine osmolality will be measured as supportive of target engagement in phase 1 studies. Positive effects on TKV, seen in vivo in the Pkd1RC/RC model, will be assessed in a phase 2 clinical trial. By directly targeting cAMP, LoAc® compounds have the potential to slow renal disease progression, and to delay or prevent the need for dialysis, offering a potential new treatment option for ADPKD patients.

Investigation of heat stress responses and adaptation mechanisms by integrative metabolome and transcriptome analysis in tea plants (Camellia sinensis).

Extreme high temperature has deleterious impact on the yield and quality of tea production, which has aroused the attention of growers and breeders. However, the mechanisms by which tea plant varieties respond to extreme environmental heat is not clear. In this study, we analyzed physiological indices, metabolites and transcriptome differences in three different heat-tolerant tea plant F1 hybrid progenies. Results showed that the antioxidant enzyme activity, proline, and malondialdehyde were significantly decreased in heat-sensitive 'FWS' variety, and the accumulation of reactive oxygen molecules such as H2O2 and O2- was remarkably increased during heat stress. Metabolomic analysis was used to investigate the metabolite accumulation pattern of different varieties in response to heat stress. The result showed that a total of 810 metabolites were identified and more than 300 metabolites were differentially accumulated. Transcriptional profiling of three tea varieties found that such genes encoding proteins with chaperon domains were preferentially expressed in heat-tolerant varieties under heat stress, including universal stress protein (USP32, USP-like), chaperonin-like protein 2 (CLP2), small heat shock protein (HSP18.1), and late embryogenesis abundant protein (LEA5). Combining metabolomic with transcriptomic analyses discovered that the flavonoids biosynthesis pathway was affected by heat stress and most flavonols were up-regulated in heat-tolerant varieties, which owe to the preferential expression of key FLS genes controlling flavonol biosynthesis. Take together, molecular chaperons, or chaperon-like proteins, flavonols accumulation collaboratively contributed to the heat stress adaptation in tea plant. The present study elucidated the differences in metabolite accumulation and gene expression patterns among three different heat-tolerant tea varieties under extreme ambient high temperatures, which helps to reveal the regulatory mechanisms of tea plant adaptation to heat stress, and provides a reference for the breeding of heat-tolerant tea plant varieties.

Development and application of KASP marker for high throughput detection of the seedless trait in grapevine

Molecular marker-assisted selection (MAS) can significantly accelerate and improve the efficiency of the breeding process in seedless grape cultivars. In this study, we developed the KASP_VviAGL11 and VviAGL11_410 markers based on a single nucleotide polymorphism (SNP) site (Chr18: 26889437 (A/C)) of the VviAGL11 gene, and compared them with previously reported SSR markers p3_VvAGL11 and 5U_VviAGL11 by testing 101 cultivars and 81 F1 hybrid progenies. The results showed that both of the proposed markers obtained 100% accuracy rates in detecting allele A, which was closely associated with the seedless trait in grapes, while p3_VvAGL11 and 5U_VviAGL11 had lower accuracy rates due to their tendency to produce false positives. After careful evaluation of the technical advantages and disadvantages associated with these markers, we concluded that KASP_VviAGL11 was superior in terms of simplicity, cost-effectiveness, efficiency, and accuracy. Thus, we optimized the process of molecular MAS for seedless grapes, focusing on the KASP_VviAGL11 marker as a central component, to provide key technical support for the development of new seedless grape cultivars.

Heterosis and Mixed Genetic Analysis of Flowering Traits in Cross Breeding of Day-Neutral Chrysanthemum (Asteraceae)

Day-neutral multiflora chrysanthemums can flower throughout the year without being influenced by daylength and have great application value in gardens. Studying heterosis and the genetic basis of important traits in day-neutral chrysanthemums can accelerate the breeding of new cultivars. In this research, a genetic population was constructed by crossing 135 F1 hybrid progeny from the day-neutral chrysanthemum ‘82-81-19’ (female parent) and the late-flowering chrysanthemum ‘388Q-76’ (male parent). Six traits, including abnormal (crown) bud, plant height, plant crown width, budding date, full flowering date, and number of petal layers, were selected for inheritance and heterosis analyses, and a single-generation major gene plus polygene mixed inheritance model was used to perform mixed inheritance analysis on these traits. The results indicated that the six traits were widely segregated in the F1 population, with the coefficient of variation (CV) ranging from 30% to 84%. The phenomena of heterosis and extra-parent segregation existed generally in F1 progeny, and the ratio of heterosis value of mid-parents (RHm) for the six traits was 45.5%, 2%, 2%, 6%, 6%, and −0.3%, respectively. The mixed genetic analysis showed that the abnormal (crown) bud and budding date were fitted to the B-3 model and controlled by two pairs of additive major genes. The plant height and plant crown width were fitted to the A-0 model, and no major gene was detected. The full flowering date was fitted to the A-1 model and was controlled by one pair of major genes. The number of petal layers was fitted to the B-1 model and controlled by two pairs of additive–dominant major genes. The heritabilities of major genes for abnormal bud, budding date, full flowering date, and the number of petal layers were 1.0, 0.9871, 0.7240, and 0.5612, respectively, indicating that these traits were less affected by environmental factors. Using a percentile scoring method, eight day-neutral chrysanthemum genotypes were selected from the hybrid progeny.

Evaluation of intergeneric F1 hybrid progenies of papaya (Arka Prabhath x Vasconcellea cauliflora and Arka Prabhath x Vasconcellea cundinamarcencis) for morphological, fruit and yield traits coupled with PRSV tolerance

Papaya is one of the most important fruit crops of tropical and subtropical regions of the world including India. Though India stands first in production in the world, the productivity is low as compared to other countries due to high incidence of papaya ring spot virus (PRSV-P) attack. As all the cultivated varieties under genus Carica are susceptible to PRSV, investigations were carried out to evaluate fifteen intergeneric hybrid progenies of Arka Prabhath x V. cauliflora and eighty-five progenies of Arka Prabhath x V. cundinamarcencis for morphological, fruit and yield traits coupled with PRSV- P tolerance. Out of fifteen, seven progenies of ‘Arka Prabhath’ x V. cauliflora viz., IGHF1S4-1, IGHF1S4-12, IGHF1S4-13, IGHF1S4-14, IGHF1S4-15, IGHF1S4-17, IGHF1S4-18 and out of eighty-five, six progenies of ‘Arka Prabhath’ x V. cundinamarcensis viz., IGHF1S1-17, IGHF1S1-19, IGHF1S6-20, IGHF1S2-14, IGHF1S5-12 and IGHF1S5-14 recorded desirable traits such as days to first fruiting (240 to 250 days), bearing height (48 to 74 cm), plant height (175 to 200 cm), trunk circumference (37 to 48 cm), fruit weight (1133.67 to 2202.00 g), pulp thickness (2.45 to 4.05 cm), TSS (11.50 to 13.80 ºBrix), fruits/tree (40 to 58) and yield (45.00 to 78.20 kg/tree) coupled with PRSV tolerance with disease score1 (only a few tiny chlorotic spots on leaves). These progenies were selected and forwarded for next generation (F2). The hybridity was also confirmed using SSR marker (mCpCIR59).

Forming innovative clonal rootstocks for varieties of the genus <i>Cerasus</i> Mill. (<i>Prunus cerasus</i> L.)

The paper considers the main problems and solutions reported in literature concerning the formation and use of clonal rootstocks for cherry varieties in Russia, Europe and America. Examples of certain species are given (Cerasus fruticosa Pall., C.incisa Thunb., C.canescens (Bois) Erem. et Yushev, C.pseudocerasus Fr. Schmidt.), which were previously involved in hybridization for forming clonal rootstock of stone fruit crop. Following the performed research, the authors propose an approach to forming innovative vegetatively propagated rootstocks for species of the genus Cerasus Mill. (cherry, sweet cherry, Oriental cherry) based on distant hybridization and using germ preselection and cultivation in vitro. The obtained results confirm the importance of including previously selected sources and donors in the breeding process for forming clonal rootstocks with the following essential features: resistance to adverse environmental factors, low and high air temperatures, main harmful diseases – coccomycosis (causative agent Coccomyces hiemalis Higg.), monilial burn (Monilia cinerea Bon.), clasterosporiosis (Clasterosporium carpophilum Aderch.), and easy rooting of shoots (rooting ability). Such sources were isolated from collection representatives of cherry species: C.sachalinensis (Fr. Schm.) Komar. et Klob.-Alis. – 49 specimens, C.serrulata (Lindl.) G. Don. – 35, C.kurilensis (Miyabe) Kaban et Vorobiev – 43, (C.maximowiczii (Rupr.) Komar) – 34, C.lannesiana Carr – 4. The above forms, selected for breeding traits for the rootstock, were used for hybridization. Forms were isolated from F1 hybrid progeny from crossing them with sweet cherry (both forward and backward), and 25 F2 hybrids were obtained from all representatives. Germ cultivation in vitro enabled the breeding process to be accelerated by 2–3 years, and micropropagation of hybrid clones in vitro, transfer of microplants ex vitro and their subsequent cultivation in vivo under laboratory conditions enabled the necessary amount of breeding plant material (75 hybrids and 650 clones) to be obtained for further tests in a nursery.

Identification of F1 Hybrid Progenies in Mango Based on Fluorescent SSR Markers

Mango (Mangifera indica L.) belongs to the genus Mangifera and family Anacardiaceae, and is an important tropical fruits. Artificial cross breeding (hand pollination) is an important method for breeding new mango cultivars. It is easy to produce false hybrids in the process of artificial pollination breeding. Therefore, it is necessary to establish rapid and accurate molecular detection methods to identify the authenticity of hybrids. Mango ‘Jinhuang’ and ‘Renong No.1′ and 65 individual plants of their F1 hybrids were used as experimental materials, eight SSRs (simple sequence repeats) primer pairs with polymorphism in parents were used to identify the F1 hybrids by capillary electrophoresis. The results showed that PCR product size (bp) for eight primers ranged from 108 bp (ES55) to 176 bp (ES63) in 65 samples. A total of 62 true hybrids were identified from 65 hybrid progenies, and the true hybrid rate was 95.38%. A total of 18 alleles were amplified by eight SSRs, seven SSR loci showed binary segregations, whereas only one SSR locus ES83 showing ab:ac:bb:bc segregation fitted to the expected segregation ratio of 1:1:1:1. The value of expected heterozygosity (He), ranged from 0.34 to 0.62, whereas the value of observed heterozygosity ranged from 0.44 to 0.81. Chi-square test showed that eight SSR loci were in accordance with Mendel’s segregation law. The results of cluster analysis showed that the parents and 62 true hybrids could be classified into two categories at 0.58: the first category contained 27 offspring, clustered with ‘Jinhuang’ and showed a maternal genetic tendency. The second category contained 35 offspring, clustered with ‘Renong No.1′ and showed a partial paternal genetic tendency. DNA fingerprint of hybrids from ‘Jinhuang’ × ‘Renong No.1′ cross were constructed using eight SSR primers for variety protection.

Genetic exchange with an outcrossing sister species causes severe genome-wide dysregulation in a selfing Caenorhabditis nematode.

Different modes of reproduction evolve rapidly, with important consequences for genome composition. Selfing species often occupy a similar niche as their outcrossing sister species with which they are able to mate and produce viable hybrid progeny, raising the question of how they maintain genomic identity. Here, we investigate this issue by using the nematode Caenorhabditis briggsae, which reproduces as a hermaphrodite, and its outcrossing sister species Caenorhabditis nigoni We hypothesize that selfing species might develop some barriers to prevent gene intrusions through gene regulation. We therefore examined gene regulation in the hybrid F2 embryos resulting from reciprocal backcrosses between F1 hybrid progeny and C. nigoni or C. briggsae F2 hybrid embryos with ∼75% of their genome derived from C. briggsae (termed as bB2) were inviable, whereas those with ∼75% of their genome derived from C. nigoni (termed as nB2) were viable. Misregulation of transposable elements, coding genes, and small regulatory RNAs was more widespread in the bB2 compared with the nB2 hybrids, which is a plausible explanation for the differential phenotypes between the two hybrids. Our results show that regulation of the C. briggsae genome is strongly affected by genetic exchanges with its outcrossing sister species, C. nigoni, whereas regulation of the C. nigoni genome is more robust on genetic exchange with C. briggsae The results provide new insights into how selfing species might maintain their identity despite genetic exchanges with closely related outcrossing species.

Metabolomic and transcriptomic analyses reveal a MYB gene, CsAN1, involved in anthocyanins accumulation separation in F1 between 'Zijuan' (Camellia sinensis var. assamica) and 'Fudingdabaicha' (C. sinensis var. sinensis) tea plants.

‘Zijuan’ (Camellia sinensis var. assamica), a somatic mutant with purple foliage and stem selected from the Yunnan Daye cultivar, has been well developed owing to abnormal pattern of anthocyanin accumulation. However, the genetic basis for the specific accumulation of phloem glycosides is not clear. Tea plants are self-incompatible, so parents with large differences in foliage color were used for crosses to investigate the genetic mechanism of anthocyanins. In this study, ‘Zijuan’ and green foliage cultivar ‘Fudingdabaicha’ (C. sinensis var. sinensis) were used as female and male parents, respectively, to generated F1 hybrid progenies with various anthocyanin contents. In order to decipher the genetic rules of anthocyanins accumulation, we performed widely targeted metabolic and transcriptomic profiling. The results showed that cyanidin-3-O-galactoside, delphinidin-3-O-galactoside and petunidin-3-O-galactoside were the major types of anthocyanins and factors directly led to the color variation between parents and F1 plants. Transcriptomic analyses suggested the significant up-regulation of anthocyanidin synthase gene (CsANS1) and CsAN1, a MYB family gene positively regulated the expression of CsANS1, in anthocyanin-rich tea plants. Furthermore, the deletion mutation of CsAN1 was found by cloning and alignment in anthocyanin-lacking cultivars. Taken together, the function deficiency of CsAN1 is predominantly responsible for the inability of anthocyanins accumulation, and this trait is heritable in progenies through hybridization. The present study elucidated the molecular basis of leaf purple trait formation in ‘zijuan’ and ‘Fudingdabaicha’ and their F1 plants, which helps to elucidate the genetic mechanism of leaf anthocyanin accumulation regulation in tea plants, and the results provide a research reference for the selection and breeding of high anthocyanin type tea varieties.

Characteristics and Fitness Analysis through Interspecific Hybrid Progenies of Transgenic Brassica napus and B. rapa L. ssp.

Interspecific hybridization between transgenic crops and their wild relatives is a major concern for transgene dispersal in the environment. Under controlled conditions, artificial hand pollination experiments were performed in order to assess the hybridization potential and the fitness of interspecific hybrids between Brassica rapa and genetically modified (GM) Brassica napus. Initially, six subspecies of B. rapa were hybridized with GM B. napus through hand pollination. In the resulting F1 hybrids, the combination of B. rapa ssp. narinosa (♀) × GM B. napus (♂) had the highest crossability index (16.9 ± 2.6). However, the F1 selfing progenies of B. rapa ssp. rapa (♀) × GM B. napus were found to be more effective in producing viable future generations with the highest crossability index (1.6 ± 0.69) compared to other subspecies. Consequently, they were used for the generation of F2 and F3 progenies. The 18 different morphological characteristics among the parental cross-combinations and F1 hybrid progenies were measured and visualized through hierarchical clustering. Different generations were found to be grouped based on their different morphological characteristics. The chromosome numbers among the interspecific hybrids ranged from 2n = 29 to 2n = 40. Furthermore, the SSR markers revealed the presence of genomic portions in the hybrids in comparison with their parental lines. There is a high possibility of transgene flow between GM B. napus and B. rapa. The study concluded that the interspecific hybrids between B. napus and B. rapa can be viable and can actively hybridize up to F3 generations and more. This suggests that the GM B. napus can disperse the transgene into B. rapa, and that it can pass through for several generations by hand pollination in a greenhouse environment.

Identification and genetic diversity analysis of hybrid offspring of azalea based on EST-SSR markers

Azalea is a world famous flower with high economic and ornamental value. The breeding of new azalea varieties is mainly done by cross breeding. However, there is a risk that cross breeding may cause errors in the hybrid offspring due to contamination by exogenous pollen. Therefore, the identification of hybrid offspring is an important part of azalea breeding. In this study, the parents of three hybrid combinations and their 88 F1 hybrid offspring were selected to screen 15 pairs of EST-SSR primers to identify the authenticity of azalea hybrid offspring. The results showed that the authenticity of 88 azalea F1 hybrid progenies could be determined by at least four primer pairs. Genetic diversity analysis of azalea hybrid progeny revealed that the number of alleles and polymorphic information content of the progeny increased to different degrees, and the more distant the genetic distance between parents, the richer the polymorphic information. It is suggested that EST-SSR molecular marker can be applied for the early identification and genetic diversity analysis of the progeny of azalea hybrids. This method is of positive significance for improving the breeding efficiency of new varieties and exploring the genetic background of azalea.

Screening and Validation of SSR Molecular Markers for Identification of Downy Mildew Resistance in Intraspecific Hybrid F1 Progeny (V. vinifera)

Downy mildew is a major disease that severely restricts the healthy and sustainable development of the global grape and wine industry, so there is significant interest in breeding high-quality disease-resistant varieties. In this study, hybridization was carried out between two disease-resistant and four high-quality varieties for eight hybrid combinations. The downy mildew resistance of 6 hybrid parents and 350 hybrid F1 progeny was determined by the leaf disc inoculation test, and the frequency distribution of the disease resistance grades was analyzed for the different populations. Agarose gel electrophoresis and disease resistance identification were used to screen SSR molecular markers of the tested hybrids for resistance to downy mildew. The results showed that minor disease-resistance genes in V. vinifera varieties were the main determinant of the variation in disease susceptibility among different varieties. V. vinifera resistance to downy mildew is inherited by a quantitative trait controlled by multiple genes. Intraspecific hybridization of V. vinifera can be applied to breed suitable intermediate materials or new types of disease resistance. The screening and verification of simple-sequence repeats (SSR) as molecular markers revealed that UDV-304, UDV-305, and UDV-737 could be used as standard markers for the identification of downy mildew resistance of hybrid progeny of Ecolly and Cabernet Sauvignon, UDV-305 could be used for the hybrid progeny of Meili and Cabernet Sauvignon, and VMC8g9 could be used for the hybrid progeny of Ecolly or Meili and Dunkelfelder. The disease-resistant progeny selected in this study can be used as intermediate materials for the breeding of new high-quality, disease-resistant varieties by intraspecific recurrent selection in V. vinifera. The screened standard markers can be utilized for rapid batch identification of hybrid progeny of different hybrid combinations to facilitate molecular marker-assisted breeding.

COMPARATIVE MORPHOLOGY, BIOSYSTEMATICS, AND REPRODUCTIVE ECOLOGY OF PHACELIA STEBBINSII, P. MARCESCENS, AND P. QUICKII

Phacelia stebbinsii Constance & Heckerd, P. marcescens Eastw. ex J.F.Macbr., and P. quickii J.T.Howell are morphologically similar annual taxa endemic to the west side of the Sierra Nevada in California. Experimental crosses show P. quickii is reproductively isolated from the other two species, and that P. marcescens and P. stebbinsii readily produce fully fertile F1 and S2 hybrid progeny, but typically only when P. marcescens is the carpellate parent. The presence of geographic separation, habitat differences, unilateral crossability, pollen certation, and problems with cotyledon emergence in hybrid seedlings provide evidence P. stebbinsii is distinct from P. marcescens and they are maintained as separate species. Flower and fruit characters used to distinguish these three species from each other appear to be related to differences in seed dispersal and seedling establishment in different habitats.

Molecular Characterization and Genetic Structure Evaluation of Breeding Populations of Fennel (Foeniculum vulgare Mill.)

Fennel, or Foeniculum vulgare Mill., is an important horticultural crop belonging to the Apiaceae family that is cultivated worldwide and used in the agri-food sector and for pharmaceutical preparations. Breeding strategies in this species usually involve three parental lines, including two maternal lines (one cytoplasmic male-sterile line and an ideotype representative maintainer line) that are crossed to obtain an ideotype representative of the cytoplasmic male-sterile line and one paternal line, used as a pollinator in crosses with the progeny of the derived maternal lines. From this cross, F1 hybrid progenies are obtained, which are characterized by high levels of heterozygosity and hybrid vigor. In this study, over 450 plants, representing 8 breeding populations and their respective 3 parental and 1 progeny line, were genotyped by means of codominant molecular markers. The 12 highly polymorphic microsatellites enabled the analyses of the genetic variability, distinctiveness and stability of each breeding line. Moreover, the genetic structure of the core collection was investigated, which, together with the homozygosity, gene flow and genetic similarity results, allowed the identification of unsuitable lines to be used in breeding plans due to their low homozygosity (10.4% in the pollinator line of population 7). Moreover, the Bayesian reconstruction of the core collection’s genetic structure, based on the codominant markers used, allowed us to confirm the distinctiveness results obtained from the genetic similarity investigation and the computed gene flow estimates. Among these, a trend in hybrid heterozygosity was also observed, that increased when the genetic similarity between the respective parental lines decreased. Thus, this research proposes a suitable method for genotyping fennel populations in pre- and post-breeding approaches, such as marker-assisted breeding or breeding line distinctiveness and stability verifications.

Inheritance of heat tolerance in perennial ryegrass (Lolium perenne, Poaceae): evidence from progeny array analysis.

BackgroundHeat stress is considered one of the most important environmental factors influencing plant physiology, growth, development, and reproductive output. The occurrence and damage caused by heat stress will likely increase with global climate change. Thus, there is an urgent need to better understand the genetic basis of heat tolerance, especially in cool season plants.Materials and MethodsIn this study, we assessed the inheritance of heat tolerance in perennial ryegrass (Lolium perenne L. subspecies perenne) , a cool season grass, through a comparison of two parental cultivars with their offspring. We crossed plants of a heat tolerant cultivar (Kangaroo Valley) with plants of a heat sensitive cultivar (Norlea), to generate 72 F1 hybrid progeny arrays. Both parents and their progeny were then exposed to heat stress for 40 days, and their photosynthetic performance (Fv/Fm values) and leaf H2O2 content were measured.ResultsAs expected, Kangaroo Valley had significantly higher Fv/Fm values and significantly lower H2O2 concentrations than Norlea. For the F1 progeny arrays, values of Fv/Fm decreased gradually with increasing exposure to heat stress, while the content of H2O 2 increased. The progeny had a wide distribution of Fv/Fm and H 2O2 values at 40 days of heat stress. Approximately 95% of the 72 F1 progeny arrays had Fv/Fm values that were equal to or intermediate to the values of the two parental cultivars and 68% of the progeny arrays had H2O2 concentrations equal to or intermediate to their two parents.ConclusionResults of this study indicate considerable additive genetic variation for heat tolerance among the 72 progeny arrays generated from these crosses, and such diversity can be used to improve heat tolerance in perennial ryegrass cultivars. Our findings point to the benefits of combining physiological measurements within a genetic framework to assess the inheritance of heat tolerance, a complex plant response.

Detection and diversity of viruses infecting African yam (Dioscorea rotundata) in a collection and F1 progenies in Côte d'Ivoire shed light to plant-to-plant viral transmission.

Yam (Dioscorea spp.) is a major staple food whose production is hampered by viral diseases. However, the prevalence, diversity, transmission, and impact of yam‐infecting viruses remain poorly documented. This study reports on the symptomatology, prevalence, and molecular diversity of eight viruses in 38 D. rotundata accessions from a germplasm collection and 206 F1 hybrid progenies maintained in Côte d'Ivoire. Mean severity scores as assessed from leaf symptoms ranged from 2 to 4 in the germplasm collection and from 1 to 3 in F1 hybrids, respectively. Dioscorea mosaic‐associated virus (DMaV), potexviruses, and yam mosaic virus (YMV) were detected by PCR‐based diagnosis tools in single and mixed infections in both the D. rotundata collection and F1 progenies, whereas badnaviruses were detected only in the germplasm collection. In contrast, cucumber mosaic virus (CMV), yam macluraviruses, yam asymptomatic virus 1 (YaV1), and yam mild mosaic virus (YMMV) could not be detected. No correlation could be established between severity scores and indexing results. Phylogenetic analysis performed on partial viral sequences amplified from infected samples unveiled the presence of two putative novel viral species belonging to genera Badnavirus and Potexvirus and provided evidence for plant‐to‐plant transmission of YMV, DMaV, and yam potexviruses.

Profiling of Volatile Compounds and Associated Gene Expression in Two Anthurium Cultivars and Their F1 Hybrid Progenies.

Anthurium is an important ornamental crop in the world market and its floral scent can enhance its ornamental value. To date, studies of the components and formation mechanism of the floral scent of Anthurium are relatively few. In this study, the scent profiles of two Anthurium varieties were measured by gas chromatograph-mass spectrometer (GC-MS). There were 32 volatile organic compounds (VOCs) identified in Anthurium ‘Mystral’, and the most abundant compound was eucalyptol (57.5%). Extremely small amounts of VOCs were detected in Anthurium ‘Alabama’. Compared with A. ‘Alabama’, most genes related to floral scent synthesis exhibited a higher expression in A. ‘Mystral’, including AaDXS, AaDXR, AaMDS, AaHDS, AaTPS, AaDAHPS, AaADT2, AaPAL1, and AaPAL2. In order to produce new varieties of Anthurium with fragrance, 454 progenies of two crossbred combinations of A. ‘Mystral’ and A. ‘Alabama’ were obtained. Four F1 generation plants with different floral scent intensities were selected for further study. The major components of floral scent in the progenies were similar to that of the parental A. ‘Mystral’ plant. The expression patterns of genes related to floral scent synthesis were consistent with the relative contents of different types of VOCs. This study revealed the profiles of volatile compounds and associated gene expression in two Anthurium cultivars and their F1 hybrids, which provided a basis for the floral scent inheritance of Anthurium andraeanum.

DNA Methylation Silences Exogenous Gene Expression in Transgenic Birch Progeny.

The genetic stability of exogenous genes in the progeny of transgenic trees is extremely important in forest breeding; however, it remains largely unclear. We selected transgenic birch (Betula platyphylla) and its hybrid F1 progeny to investigate the expression stability and silencing mechanism of exogenous genes. We found that the exogenous genes of transgenic birch could be transmitted to their offspring through sexual reproduction. The exogenous genes were segregated during genetic transmission. The hybrid progeny of transgenic birch WT1×TP22 (184) and WT1×TP23 (212) showed higher Bgt expression and greater insect resistance than their parents. However, the hybrid progeny of transgenic birch TP23×TP49 (196) showed much lower Bgt expression, which was only 13.5% of the expression in its parents. To elucidate the mechanism underlying the variation in gene expression between the parents and progeny, we analyzed the methylation rates of Bgt in its promoter and coding regions. The hybrid progeny with normally expressed exogenous genes showed much lower methylation rates (0–29%) than the hybrid progeny with silenced exogenous genes (32.35–45.95%). These results suggest that transgene silencing in the progeny is mainly due to DNA methylation at cytosine residues. We further demonstrated that methylation in the promoter region, rather than in the coding region, leads to gene silencing. We also investigated the relative expression levels of three methyltransferase genes: BpCMT, BpDRM, and BpMET. The transgenic birch line 196 with a silenced Gus gene showed, respectively, 2.54, 9.92, and 4.54 times higher expression levels of BpCMT, BpDRM, and BpMET than its parents. These trends are consistent with and corroborate the high methylation levels of exogenous genes in the transgenic birch line 196. Therefore, our study suggests that DNA methylation in the promoter region leads to silencing of exogenous genes in transgenic progeny of birch.

Genetic parameters of drought tolerance for agromorphological traits in eggplant, wild relatives, and interspecific hybrids

AbstractCrop wild relatives (CWRs) have barely been used in eggplant (Solanum melongena) breeding. However, introgression breeding may help in coping with the challenges posed by climate change. During the rainy and dry seasons, we evaluated nine accessions of eggplant, nine accessions of seven related species (of which six are CWRs and one is a cultivated relative), and 12 interspecific hybrids. Drought tolerance scores, calculated as ratios of growing rates and ratios of mean values during the rainy and dry seasons, were obtained for vegetative growth and yield‐related traits. Considering the vegetative growth and yield‐related traits evaluated, the F1 hybrid progenies of eggplant with S. insanum MEL1 × INS2 and MEL4 × INS3, S. anguivi MEL6 × ANG1, and S. dasyphyllum MEL6 × DAS1, plus the S. sysimbriifolium accession SIS1, displayed drought tolerance. Small to large gaps were observed between phenotypic and genotypic CVs of drought tolerance scores, reflecting variable influence of environmental factors on drought tolerance of the traits evaluated. In addition, the narrow‐sense heritability was generally moderate, indicating that genes with dominance and/or epistasis effects may be involved in the expression of drought tolerance. High genotypic correlations for drought tolerance scores between pairs of traits such as branching index and leaf length, leaf petiole diameter and plant height, number of stamens per flower, and fruit pedicel length suggest that drought tolerance of these traits is controlled by genes that are in linkage disequilibrium and/or have pleiotropic effects. The results are relevant for the development of drought‐tolerant cultivars of eggplant.

Heritable genetic background alters survival and phenotype of Mll-AF9-induced leukemias

The MLL-AF9 fusion protein occurring as a result of t(9;11) translocation gives rise to pediatric and adult acute leukemias of distinct lineages, including acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), and mixed-phenotype acute leukemia (MPAL). The mechanisms underlying how this same fusion protein results in diverse leukemia phenotypes among different individuals are not well understood. Given emerging evidence from genome-wide association studies that genetic risk factors contribute to MLL-rearranged leukemogenesis, here we tested the impact of genetic background on survival and phenotype of a well-characterized Mll-AF9 knockin mouse model. We crossed this model with five distinct inbred strains (129, A/J, C57BL/6, NOD, CAST) and tested their F1 hybrid progeny for dominant genetic effects on Mll-AF9 phenotypes. We discovered that genetic background altered peripheral blood composition, with Mll-AF9 CAST F1 having a significantly increased B-lymphocyte frequency, while the remainder of the strains exhibited myeloid-biased hematopoiesis, similar to the parental line. Genetic background also had an impact on overall survival, with Mll-AF9 A/J F1 and Mll-AF9 129 F1 having significantly shorter survival and Mll-AF9 CAST F1 having longer survival, compared with the parental line. Furthermore, we observed a range of hematologic malignancies, with Mll-AF9 A/J F1, Mll-AF9 129 F1, and Mll-AF9 B6 F1 developing exclusively myeloid cell malignancies (myeloproliferative disorder [MPD] and AML), whereas a subset of Mll-AF9 NOD F1 developed MPAL and Mll-AF9 CAST F1 developed ALL. This study provides a novel in vivo experimental model in which to evaluate the underlying mechanisms by which MLL-AF9 results in diverse leukemia phenotypes and provides definitive experimental evidence that genetic risk factors contribute to survival and phenotype of MLL-rearranged leukemogenesis.