Parental Lines Research Articles

Microcystin-LR-induced epithelial-mesenchymal transition-like cells acquire resistance to multi-toxins

The protein phosphatase inhibitor microcystin-LR (MC-LR), a hepatocyte-selective cyanotoxin, induces phenotypic changes in HEK293 OATP1B3-expressing (HEK293-OATP1B3) cells, which include cytoskeletal reorganization (HEK293-OATP1B3-AD) and anoikis resistance (HEK293-OATP1B3-FL) transformed cells, respectively. These cells acquire resistance to MC-LR and partial epithelial-mesenchymal transition (EMT) characteristics. In cancer cells, EMT is generally involved in multi-drug resistance. Here, we focused on the multi-drug resistance of HEK293-OATP1B3-AD and HEK293-OATP1B3-FL cells. The MTT assay and immunoblotting were conducted to examine the responses of HEK293-OATP1B3, HEK293-OATP1B3-AD, and HEK293-OATP1B3-FL cells to multiple toxins and drugs that function as substrates for OATP1B3, including MC-LR, nodularin (Nod), okadaic acid (OA), and cisplatin (CDDP). HEK293-OATP1B3-AD and HEK293-OATP1B3-FL cells were more resistant to MC-LR, Nod, and OA than HEK293-OATP1B3 cells. Conversely, the three cell types were equivalently sensitive to CDDP. By using protein phosphatase assay, the reduction of the inhibitory effect of MC-LR and Nod on phosphatase activity might be one reason for the resistance to MC-LR and Nod in HEK293-OATP1B3-AD and HEK293-OATP1B3-FL cells. Furthermore, the parental HEK293-OATP1B3 cells showed enhanced p53 phosphorylation and stabilization after MC-LR exposure, while p53 phosphorylation was attenuated in HEK293-OATP1B3-AD and HEK293-OATP1B3-FL cells. Moreover, in HEK293-OATP1B3-AD and HEK293-OATP1B3-FL cells, AKT phosphorylation was higher than that of the parental HEK293-OATP1B3 cell line. These results suggest that the multi-toxin resistance observed in HEK293-OATP1B3-AD and HEK293-OATP1B3-FL cells is associated with AKT activation and p53 inactivation.

CA-CAS-01-A: A Permissive Cell Line for Isolation and Live Attenuated Vaccine Development Against African Swine Fever Virus

African swine fever virus (ASFV) is the causative agent of the highly lethal African swine fever disease that affects domestic pigs and wild boars. In spite of the rapid spread of the virus worldwide, there is no licensed vaccine available. The lack of a suitable cell line for ASFV propagation hinders the development of a safe and effective vaccine. For ASFV propagation, primary swine macrophages and monocytes have been widely studied. However, obtaining these cells can be time-consuming and expensive, making them unsuitable for mass vaccine production. The goal of this study was to validate the suitability of novel CA-CAS-01-A (CAS-01) cells, which was identified as a highly permissive cell clone for ASFV replication in the MA-104 parental cell line for live attenuated vaccine development. Through a screening experiment, maximum ASFV replication was observed in the CAS-01 cell compared to other sub-clones of MA-104 with 14.89 and log10 7.5 ± 0.15 Ct value and TCID50/ml value respectively. When CAS-01 cells are inoculated with ASFV, replication of ASFV was confirmed by Ct value for ASFV DNA, HAD50/ml assay, TCID50/ml assay, and cytopathic effects and hemadsoption were observed similar to those in primary porcine alveolar macrophages after 5th passage. Additionally, we demonstrated stable replication and adaptation of ASFV over the serial passage. These results suggest that CAS-01 cells will be a valuable and promising cell line for ASFV isolation, replication, and development of live attenuated vaccines.

Gene Flow Between the Wild Rice Species (Oryza longistaminata) and Two Varieties of Cultivated Rice (Oryza sativa) in Kilombero District, Tanzania

The direction and rate of gene flow between the perennial wild rice species, O. longistaminata and cultivated rice, O. sativa was investigated using nine Microsatellite (SSR) markers. The study involved characterization of parental lines of two cultivated rice varieties and O. longistaminata and their F1 progenies. Presence of crop-specific alleles in the wild species or vice-versa was used as an indicator of occurrence of gene flow between the two species. The study revealed that gene flow between O. longistaminata and each of the two varieties of O. sativa occurs naturally. The direction of gene flow was mainly from the cultivated to wild species. The rate (frequency) of gene flow was higher from cultivated (O. sativa) to wild rice (O. longistaminata) than from O. longistaminata to O. sativa and varied with cultivated rice variety. Higher rate of gene flow from cultivated to wild species could be due to floral structure and out-crossing nature of the wild species (O. longistaminata). Gene flow between O. longistaminata and O. sativa is likely to change genetic integrity of natural populations of the two species in areas where they occur sympatrically. However, isolation by distance can help to control gene flow between these species.

Comparative and integrative omic analysis focused on chaperones and interactors in a cultivated and an exotic tomato at different fruit ripening stages

Heat Shock Proteins (HSPs) are a superfamily of chaperones that have been characterized in different organisms. In plants, HSPs promote protein folding and deaggregation during abiotic stress or developmental changes. The aim of this work was to integrate several omic-data to identify chaperone and putative interactors in Solanum lycopersicum domesticated cultivar Caimanta (C) and in the latinoamerican wild Solanum pimpinellifolium (P) genotypes during fruit ripening (FR), which are the parental lines of different breeding populations obtained by our research group. We were able to identify newly putative interactors and simultaneously induced HSP members at the transcription and proteomic levels. This integrative approach also revealed gene/protein families related to chlorophyll content, photosynthesis and HSP70 chaperones in C. Furthermore, P was enriched with chaperones, including HSP20, ATPase families, (characteristic of HSP90 and HSP100) and other protein families involved in oxidoreductase activity, supporting the hypothesis of the existence of a relationship between HSPs and developmental processes as FR. Finally, we found that some of these up-regulated chaperones show the presence of heat shock element motifs in their promoters. Proteomic coupled with transcriptomics and interactomics facilitated the exploration of a good new gene-context at the tomato development.

Agronomical Performances of Gajah Mungkur Mutant Rice Varieties Under Drought Stress

The productivity of the rice plant is commonly influenced by both genotype and environmental factors. One common environmental factor that leads to harvest failure is drought, often caused by low rainfall. Therefore, the development of drought-tolerant varieties should be implemented to obtain optimum productivity under an unfavorable environment. One of the canonical approaches for achieving this is genetic manipulation, such as by gamma radiation-induced mutation. This study aimed to determine the drought tolerance and quantitative characteristics of mutant rice varieties generated from gamma radiation. The methodology used was a split-plot design with two factors, including drought as the first factor consisting of three groups namely control, mid-level, and high-level. The second factor was rice genotypes, which consisted of six genotypes comprising PMG 07/PsJ, PMG 08/ PsJ, and PMG 09/ PsJ (mutant varieties), Gajah Mungkur (parental background), Limboto (drought-tolerant), and IR 20 (drought-susceptible). The data were statistically examined using Analysis of Variance (ANOVA) and further analyzed with Duncan’s Multiple Range Test (DMRT) with a significance level of 5%. The results showed that high-level drought significantly affected plant height during both the vegetative and mature stages. Among the mutants, PMG 08/PsJ exhibited better vegetative growth under dry conditions, retaining a relatively higher height. Drought also had a negative impact on the number of tillers and productive tillers. The PMG 08/PsJ mutant had a slightly higher number of tillers under drought cultivations. On the other hand, PMG 09/PsJ was found to have a relatively more filled grain number per panicle. Leaf rolling and dryness index showed similar trends in all varieties, while drought treatments imposed a delay on the flowering and harvesting age. Although the results demonstrated no substantial improvement over the parental lines, the selected mutant lines provided several beneficial agronomical features such as maintenance of plant height, tiller number, and shorter life cycles under severe drought. These characteristics could serve as valuable genetic resources for breeding programs focused on developing drought-resistant rice varieties for challenging environments.

Genetic diversity among maize (Zea mays L.) inbred lines adapted to Japanese climates.

Understanding the genetic diversity of inbred lines is vital for development of superior F1 varieties. The present study aimed to analyze Japanese maize parental inbred lines and determine their genetic diversity for future breeding. Genetic analyses were conducted using multiple methods. Principal component analysis (PCA), phylogenetic trees, and Bayesian clustering reflected borders between heterotic groups according to the derivation of each inbred line. A self-pollinated line derived from a classic F1 variety and another line from an open-pollinated population from the same derivation were classified as separate components by PCA and Bayesian clustering. The result suggests that open pollination could be essential in modern breeding. Of those classified as dent or flint based on their derivation, some had a combination of all components or clusters. Therefore, the classification of inbred lines should be based on their derivation and DNA markers. The findings will be valuable for breeding and genetic studies in Japan. Additionally, these techniques may be used to obtain a more significant number of SNPs and related phenotypic data.

GENETIC EVALUATION FOR SEED YIELD AND ITS COMPONENT TRAITS IN SUNFLOWER (HELIANTHUS ANNUUS L.) USING LINE × TESTER APPROACH

Sunflower is a vital oilseed crop, but yield improvement remains a major objective. Exploring germplasm and exploiting non-additive gene action can help overcome yield limitations and identify superior parental lines and crosses for hybrid development through GCA and SCA. Four parental lines (A-1, A-2, A-3, and A-4) were crossed with three testers (B-1, B-2, B-3) in a line × tester fashion to generate F0 seeds. These seeds were planted using a randomized complete block design (RCBD) with three replicate plots in the field to ensure unbiased results. Data on various growth and yield parameters is collected and analyzed using Analysis of variance and combining ability analysis. Lines A-3 and A-4 and tester B-3 demonstrated consistently positive GCA effects for several traits, signifying their broad genetic value for hybrid development. The hybrid A-4 × B-1 emerged as the top performer, exhibiting exceptional SCA effects in days to flowering, seed yield per head and height of the plant, indicating exceptional compatibility between these parental lines. Lines A-3 and A-4 displayed significant positive GCA effects for key parameters like head diameter and seed yield. Tester B-3 also contributed to enhanced yield traits in hybrid combinations. Lines A-3 and A-4, tester B-3, and hybrid A-4 × B-1 emerged as promising parents for sunflower hybrid development due to their superior GCA and SCA effects. These findings offer valuable resources for future breeding programs that enhance sunflower yield and productivity.

Effect of Hybridization on the Nutrient Compositions of Two Cultivars of Solanum aethiopicum L. (Solanaceae) Found in Anambra State

The desire for modification of valuable crop species for human nutritional fulfilment and economic growth is essential. Two cultivars of Solanum aethiopicum L. found in Anambra State were hybridized to raise an F1 that might combine the attributes of the parental lines for enhancement in yield and biochemical contents for health nutritional requirements. The emasculation method was adopted, using Solanum aethiopicum var “Anara Adazi as the female parent and transference of functional pollen from Solanum aethiopicum var “Anara” to the stigmas of the emasculated plant at the correct time. Nutrient analysis of the fruits of F1 and the parental plants was carried out using the standard method. Analysis of variance was used for data analysis. Results revealed that parental plants and F1 contained investigated nutrients in varied amounts. Ash content, crude fiber and protein were higher in the fruit of F1 hybrid (10.90±0.11, 21.50±1.51, and 5.01±0.24) respectively when compared to the fruits of the parental plants. Data obtained indicated that when two Solanum aethiopicum plants of different but closely related cultivars are crossbred, a new hybrid that possesses enhanced characteristics of the two parent plants are produced. Results of the nutrients analysis demonstrated that hybridization could significantly increase food nutrients in eggplant (S. aethiopicum).

Parent-of-origin effects for the number of oocytes and embryos in Gir cattle

Imprinting is a phenomenon that alters the expression of genes according to the parental origin of their alleles. A quantitative form to evaluate the imprinting effect is known as parent-of-origin effect. Our aim with this work is to identify parent-of-origin effects that influence the number of oocytes and embryos in Gir dairy cattle. A dataset with 17,526 Ovum Pick Up observations from 1641 Gir donors was used to estimate parent-of-origin effects for the traits number of total oocytes (TO), number of viable oocytes (VO) and number of embryos (EM). To identify parent-of-origin effects, dam and sire gametic effects were included, individually or together, in an animal model for TO, VO and EM traits. For TO, inclusion of paternal origin effects in the model was significant (P < 0.05), and explained 6 % of the total phenotypic variance. For VO and EM no significant parent-of-origin effects were found for either parental line. In conclusion, paternal effects appear to influence the total oocyte production in the Gir cattle breed.

Truncated O-glycosylation in metastatic triple-negative breast cancer reveals a gene expression signature associated with extracellular matrix and proteolysis

Breast cancer (BC) is the leading cause of death by cancer in women worldwide. Triple-negative (TN) BC constitutes aggressive and highly metastatic tumors associated with shorter overall survival of patients compared to other BC subtypes. The Tn antigen, a glycoconjugated structure resulting from an incomplete O-glycosylation process, is highly expressed in different adenocarcinomas, including BC. It also favors cancer growth, immunoregulation, and metastasis in TNBC. This work describes the differentially expressed genes (DEGs) associated with BC aggressiveness and metastasis in an incomplete O-glycosylated TNBC cell model. We studied the transcriptome of a TNBC model constituted by the metastatic murine 4T1 cell line that overexpresses the Tn antigen due to a mutation in one of the steps of the O-glycosylation pathway. We analyzed and compared the results with the parental wild-type cell line and with a Tn-negative cell clone that was poorly metastatic and less aggressive than the 4T1 parental cell line. To gain insight into the generated expression data, we performed a gene set analysis. Biological processes associated with cancer development and metastasis, immune evasion, and leukocyte recruitment were highly enriched among functional terms of DEGs. Furthermore, different highly O-glycosylated protein-coding genes, such as mmp9, ecm1 and ankyrin-2, were upregulated in 4T1/Tn+ tumor cells. The altered biological processes and DEGs that promote tumor growth, invasion and immunomodulation might explain the aggressive properties of 4T1/Tn+ tumor cells. These results support the hypothesis that incomplete O-glycosylation that leads to the expression of the Tn antigen, which might regulate activity or interaction of different molecules, promotes cancer development and immunoregulation.

The Combining Ability and Heterosis Analysis of Sweet-Waxy Corn Hybrids for Yield-Related Traits and Carotenoids.

Improving sweet-waxy corn hybrids enriched in carotenoids via a hybrid breeding approach may provide an alternative cash crop for growers and provide health benefits for consumers. This study estimates the combining ability and heterosis of sweet-waxy corn hybrids for yield-related traits and carotenoids. Eight super sweet corn and three waxy corn lines were crossed to generate 24 F1 hybrids according to the North Carolina Design II scheme, and these hybrids were evaluated across two seasons of 2021/22. The results showed that both additive and non-additive genetic effects were involved in expressing the traits, but the additive genetic effect was more predominant. Most observed traits exhibited moderate to high narrow-sense heritability. Three parental lines, namely the ILS2 and ILS7 females and the ILW1 male, showed the highest positive GCA effects on yield-related traits, making them desirable for developing high-yielding hybrids. Meanwhile, five parental lines, namely the ILS3, ILS5, and ILS7 females and the ILW1 and ILW2 males, were favorable general combiners for high carotenoids. A tested hybrid, ILS2 × ILW1, was a candidate biofortified sweet-waxy corn hybrid possessing high yields and carotenoids. Heterosis and per se performance were more positively correlated with GCAsum than SCA, indicating that GCAsum can predict heterosis for improving biofortified sweet-waxy corn hybrid enriched in carotenoids. The breeding strategies of biofortified sweet-waxy corn hybrids with high yield and carotenoid content are discussed.

Genetics, genomics, and breeding of black gram [Vigna mungo (L.) Hepper

Black gram [Vigna mungo (L.) Hepper] is a highly nutritious grain legume crop, mainly grown in South and Southeast Asia, with the largest area in India, where the crop is challenged by several biotic and abiotic stresses leading to significant yield losses. Improving genetic gains to increase on-farm yields is the primary goal of black gram breeding programs. This could be achieved by developing varieties resistant to major diseases like mungbean yellow mosaic disease, urdbean leaf crinkle virus, Cercospora leaf spot, anthracnose, powdery mildew, and insect pests such as whitefly, cowpea aphids, thrips, stem flies, and bruchids. Along with increasing on-farm yields, incorporating market-preferred traits ensures the adoption of improved varieties. Black gram breeding programs rely upon a limited number of parental lines, leading to a narrow genetic base of the developed varieties. For accelerating genetic gain, there is an urgent need to include more diverse genetic material for improving traits for better adaptability and stress resistance in breeding populations. The present review summarizes the importance of black gram, the major biotic and abiotic stresses, available genetic and genomic resources, major traits for potential crop improvement, their inheritance, and the breeding approaches being used in black gram for the development of new varieties.

Proteomic and transcriptomic characterisation of FIA10, a novel murine leukemic cell line that metastasizes into the brain.

Brain metastasis leads to increased mortality and is a major site of relapse for several cancers, yet the molecular mechanisms of brain metastasis are not well understood. In this study, we established and characterized a new leukemic cell line, FIA10, that metastasizes into the central nervous system (CNS) following injection into the tail vein of syngeneic mice. Mice injected with FIA10 cells developed neurological symptoms such as loss of balance, tremor, ataxic gait and seizures, leading to death within 3 months. Histopathology coupled with PCR analysis clearly showed infiltration of leukemic FIA10 cells into the brain parenchyma of diseased mice, with little involvement of bone marrow, peripheral blood and other organs. To define pathways that contribute to CNS metastasis, global transcriptome and proteome analysis was performed on FIA10 cells and compared with that of the parental stem cell line FDCP-Mix and the related FIA18 cells, which give rise to myeloid leukemia without CNS involvement. 188 expressed genes (RNA level) and 189 proteins were upregulated (log2 ratio FIA10/FIA18 ≥ 1) and 120 mRNAs and 177 proteins were downregulated (log2 ratio FIA10/FIA18 ≤ 1) in FIA10 cells compared with FIA18 cells. Major upregulated pathways in FIA10 cells revealed by biofunctional analyses involved immune response components, adhesion molecules and enzymes implicated in extracellular matrix remodeling, opening and crossing the blood-brain barrier (BBB), molecules supporting migration within the brain parenchyma, alterations in metabolism necessary for growth within the brain microenvironment, and regulators for these functions. Downregulated RNA and protein included several tumor suppressors and DNA repair enzymes. In line with the function of FIA10 cells to specifically infiltrate the brain, FIA10 cells have acquired a phenotype that permits crossing the BBB and adapting to the brain microenvironment thereby escaping immune surveillance. These data and our model system FIA10 will be valuable resources to study the occurrence of brain metastases and may help in the development of potential therapies against brain invasion.

Genetic assessment and potence ratio of various traits of okra (Abelmoschus esculentus) in mid hills of Himachal Pradesh

The selection of appropriate breeding strategies for achieving desired improvements in plant breeding necessitates a thorough understanding of the mechanism of gene expression implicated in the transmission of quantitative and biochemical characteristics. Line × tester study of 30 F1 hybrids obtained by crossing 13 parental lines was carried out to assess gene action and potence ratio of such traits in okra (Abelmoschus esculentus L.) during rainy (kharif) seasons of 2021 and 2022 at research farm of Dr. Yashwant Singh Parmar University of Horticulture and Forestry, Nauni, Solan, Himachal Pradesh. Out of 22 traits, 20 indicated the preponderance of non-additive gene action and additive gene action for plant height and inter-nodal length. UHFO-6 and UHFO-9 were the best general combiners, whereas, UHFO-2 × Pusa Bhindi-5 and UHFO-10 × Hisar Unnat were the best specific combiners. For 20 traits, proportional contribution of lines was higher than the testers for total variance. Potence ratio indicated that majority of the traits in the hybrids had over dominance gene effect. Over dominance was expressed for pod length, pod diameter, number of pods per plant, average pod weight, and pod yield per plant, etc. To maximize the entire genetic potential of hybrids and parents with excellent per se performance, okra crop development initiatives must include heterosis, combining ability, proportionate contribution of genotypes and potence ratio.

Interspecific hybridization and inoculation with Pantoea eucalypti improve forage performance of Lotus crop species under alkaline stress.

This study was designed to elucidate the physiological responses of three Lotus forage accessions to alkaline stress, and the influence of inoculating with Pantoea eucalypti endophyte strain on alkaline stress mitigation. A diploid L. corniculatus (Lc) accession, L. tenuis (Lt), and the interspecific hybrid Lt × Lc obtained from these two parental lines were exposed to alkaline stress (pH 8.2). Both Lt and the Lt × Lc hybrid are alkaline-tolerant compared to Lc, based on observations that dry mass was not reduced under stress, and there were no chlorosis symptoms on leaf blades. In all three Lotus accessions, Fe2+ concentration under stress decreased in aerial parts and simultaneously increased in roots. Inoculation with P.eucalypti considerably increased Fe2+ content in shoots of all three Lotus forage species under alkaline treatment. Photochemical efficiency of PSII was affected in Lc accession only when exposed to alkaline treatment. However, when cultivated under alkalinity with inoculation, plants recovered and had photosynthetic parameters equivalent to those in the control treatment. Together, the results highlight the importance of inoculation with P. eucalypti, which contributes significantly to mitigating alkaline stress. All results provide useful information for improving alkaline tolerance traits of Lotus forage species and their interspecific hybrids.

Identification of heterosis and combining ability in the hybrids of male sterile and restorer sorghum [Sorghum bicolor (L.) Moench] lines.

In sorghum [Sorghum bicolor (L.) Moench], combining ability and heterosis analysis are commonly used to evaluate superior parental lines and to screen for strongly heterotic hybrids, which helps in sorghum variety selection and breeding. In this context, combining ability and heterosis analysis were assessed using 14 restorer lines and seven cytoplasmic male sterile (CMS) lines in 2019 and 2020. The analysis of variance of all cross combinations had highly significant differences for all characters studied, which indicated a wide variation across the parents, lines, testers, and crosses. Combining ability analysis showed that the general combining ability (GCA) and specific combining ability (SCA) of the different parents were differed significantly among different traits. Most combinations with high SCA also showed high GCA in their parent lines. The heritability in the narrow sense of grain weight per panicle and grain yield was relatively low, indicating that the ability of these traits to be directly inherited by offspring was weak, that they were greatly affected by the environment. The better-parent heterosis for plant height, grain weight per panicle, panicle length, and 1000-grain weight was consistent with the order of mid-parent heterosis from strong to weak. The GCA effects of two lines 10480A, 3765A and three testers 0-30R, R111, and JY15R were significant for the majority of the agronomic traits including grain yield and might be used for improving the yield of grains in sorghum as parents of excellent specific combining ability. Seven strongly heterotic F1 hybrids were screened; of these, hybrids 3765A × R111, 1102A × L2R, and 3765A × JY15R showed significant increases in seed iristectorigenin A content and will feature into the creation of new sorghum varieties rich in iristectorigenin A.

Combining ability analysis of Cucurbita moschata D. in Côte d'Ivoire and classification of promising lines based on their gca effects.

Cucurbita moschata varieties grown in Africa have very low yield. They have been neglected, and totally ignored in agricultural research programs. However, interest in their fruits, seeds, flowers and leaves is growing nowadays due to their nutritional and medicinal potentials. That growing interest has prompted plant breeders and agronomists to develop research programs for their improvement. A complete diallel cross analysis of four parental lines, Long, Zouan-H, Oval, and Soubre and their twelve F1 hybrids, was carried out in a farming environment at the University Nangui Abrogoua, Abidjan, Côte d'Ivoire. The four parental lines and the F1 hybrids were evaluated for their general performances, combining abilities, potency ratio and heterosis effects. The investigated traits included plant height, and eleven fruit- and seed-related characters. The analysis of variance showed significant differences for all traits studied. In addition, the diallel model yielded highly significant gca effects of the female parents. The gca effects of the male parents were significant for all traits except plant height, length of the fruit, width of the fruit and length of the seed. Highly significant sca effects were observed in the crosses for all the traits. Strong maternal effects were observed for the weight and diameter of the fruit, weight of the pulp, number of seeds per fruit, weight of the fresh seeds and 100-seed weight. The general predictive ratio approached the value 1 for all the traits except weight of the fresh seed and width of the dry seed. Most of the characters under this study are predominantly determined by the effects of additive genes. But, weight of the fresh seed and width of the dry seed may be controlled by non-additive genes. Mid-parent heterosis was significant for all measured traits in the crosses, except the length of the fruit. And better-parent heterosis was significant for all traits except plant height, number of fruits per plant and length of the fruit. Gene expression is described by a super-dominance for many traits, and partial dominance for some other traits in all twelve F1 hybrids. Classification of the parental lines based on the effects of their general combining ability grouped the Soubre lines as promising contributors to fruit yield. The parental lines Long and Oval formed another group likely on the basis of the small size of their fruits, the small pulps, the smaller number of fruits per plant and the large number of seeds per fruit. However, Long would be a candidate parent for the development of cultivars with longer vegetative growth. The parental line Zouan-H formed the third group and it was mostly characterized by its large number of seeds per fruit and relatively large fruits.

Heterozigosidade, adaptabilidade e estabilidade fenotípica de genótipos de sorgo sacarino

ABSTRACT Sweet sorghum (Sorghum bicolor L.) is a promising energy crop for bioethanol production. This study aimed to investigate the influence of genetic structure on the adaptability and phenotypic stability of sweet sorghum lines and hybrids regarding the main agro-industrial traits, as well as selecting hybrids that associate high ethanol yield and yield stability in different growing environments. A total of 45 genotypes were evaluated in experiments conducted in a 5×9 triple alpha lattice design in three locations in the state of Minas Gerais, Brazil. The following traits were measured: fresh matter production, juice extraction percentage, total soluble solids content, tons of Brix per hectare, and ethanol production. Adaptability and phenotypic stability were assessed by the Wricke and Annicchiarico methods. Sweet sorghum adaptability and phenotypic stability depend on genotype genetic structure, with hybrids being more stable than parental lines. Additionally, H2x9 and H3x8 were the most promising hybrids.

Glypican-3 deficiency in liver cancer upregulates MAPK/ERK pathway but decreases cell proliferation.

Glypican-3 (GPC3) is overexpressed in hepatocellular carcinomas and hepatoblastomas and represents an important therapeutic target but the biologic importance of GPC3 in liver cancer is unclear. To date, there are limited data characterizing the biological implications of GPC3 knockout (KO) in liver cancers that intrinsically express this target. Here, we report on the development and characterization of GPC3-KO liver cancer cell lines and compare to them to parental lines. GPC3-KO variants were established in HepG2 and Hep3B liver cancer cell lines using a lentivirus-mediated CRISPR/Cas9 system. We assessed the effects of GPC3 deficiency on oncogenic properties in vitro and in murine xenograft models. Downstream cellular signaling pathway changes induced by GPC3 deficiency were examined by RNAseq and western blot. To confirm the usefulness of the models for GPC3-targeted drug development, we evaluated the target engagement of a GPC3-selective antibody, GC33, conjugated to the positron-emitting zirconium-89 (89Zr) in subcutaneous murine xenografts of wild type (WT) and KO liver cancer cell lines. Deletion of GPC3 significantly reduced liver cancer cell proliferation, migration, and invasion compared to the parental cell lines. Additionally, the tumor growth of GPC3-KO liver cancer xenografts was significantly slower compared with control xenografts. RNA sequencing analysis also showed GPC3-KO resulted in a reduction in the expression of genes associated with cell cycle regulation, invasion, and migration. Specifically, we observed the downregulation of components in the AKT/NFκB/WNT signaling pathways and of molecules related to cell cycle regulation with GPC3-KO. In contrast, pMAPK/ERK1/2 was upregulated, suggesting an adaptive compensatory response. KO lines demonstrated increased sensitivity to ERK (GDC09994), while AKT (MK2206) inhibition was more effective in WT lines. Using antibody-based positron emission tomography (immunoPET) imaging, we confirmed that 89Zr-GC33 accumulated exclusively in GPC3-expression xenografts but not in GPC3-KO xenografts with high tumor uptake and tumor-to-liver signal ratio. We show that GPC3-KO liver cancer cell lines exhibit decreased tumorigenicity and altered signaling pathways, including upregulated pMAPK/ERK1/2, compared to parental lines. Furthermore, we successfully distinguished between GPC3+ and GPC3- tumors using the GPC3-targeted immunoPET imaging agent, demonstrating the potential utility of these cell lines in facilitating GPC3-selective drug development.

Marker-assisted breeding accelerates the development of multiple-stress-tolerant rice genotypes adapted to wider environments.

Rice, one of the major staple food crops is frequently affected by various biotic/abiotic stresses including drought, salinity, submergence, heat, Bacterial leaf blight, Brown plant hopper, Gall midge, Stem borer, Leaf folder etc. Sustained increase of yield growth is highly necessary to meet the projected demand in rice production during the year 2050. Hence, development of high yielding and multiple stress tolerant rice varieties adapted to wider environments will serve the need. A systematic MAB approach was followed to pyramid eight major QTLs/genes controlling tolerance to major abiotic/biotic stresses viz., drought (qDTY1.1 and qDTY2.1), salinity (Saltol), submergence (Sub1), bacterial leaf blight (xa13 and Xa21), blast (Pi9) and gall midge (Gm4) in the genetic background of an elite rice culture CBMAS 14065 possessing high yield and desirable grain quality traits. Two advanced backcross derivatives of CBMAS 14065 possessing different combinations of target QTLs namely #27-1-39 (qDTY1.1+qDTY2.1+Sub1+xa13+Xa21+Gm4+Pi9) and #29-2-2 (qDTY1.1+qDTY2.1+Saltol+Xa21+Gm4+Pi9) were inter-mated. Inter-mated F1 progenies harboring all the eight target QTLs/genes were identified through foreground selection. Genotyping of the inter-mated F4 population identified 14 progenies possessing all eight target QTLs/genes under homozygous conditions. All the fourteen progenies were forwarded up to F8 generation and evaluated for their yield and tolerance to dehydration, salinity, submergence, blast and bacterial leaf blight. All the 14 progenies exhibited enhanced tolerance to dehydration and salinity stresses by registering lesser reduction in their chlorophyll content, relative water content, root length, root biomass etc., against their recurrent parent Improved White Ponni/CBMAS 14065. All the 14 progenies harboring Sub1 loci from FR13A exhibited enhanced survival (90 - 95%) under 2 weeks of submergence /flooding when compared to their recurrent parent CBMAS 14065 which showed 100% susceptibility The inter-mated population showed a enhanced level of resistance to bacterial leaf blight (Score = 0 to 2) against blast (Score - 0) whereas the susceptible check CO 39 and the recurrent parent CBMAS 14065 recorded high level of susceptibility (Score = 7 to 9). Our study demonstrated the accelerated development of multiple stress tolerant rice genotypes through marker assisted pyramiding of target QTLs/genes using tightly linked markers. These multiple stress tolerant rice lines will serve as excellent genetic stocks for field testing/variety release and also as parental lines in future breeding programs for developing climate resilient super rice varieties.