Search Results (61)

Grass carp is an economically important cultured species in China. Triploid embryo production is widely applied in aquaculture to achieve reproductive sterility, improve somatic growth, and reduce ecological risks associated with uncontrolled breeding. In this study, a simple cold shock method for inducing triploid grass carp was developed. The triploid induction rate of 71.73 ± 5.00% was achieved by applying a cold treatment at 4 °C for 12 min, starting 2 min after artificial fertilization. Flow cytometry and karyotype analysis revealed that triploid individuals exhibited a 1.5-fold increase in DNA content compared to diploid counterparts, with a chromosomal composition of 3n = 72 (33m + 36sm + 3st). Additionally, embryonic transcriptome analysis demonstrated that, in the cold shock-induced embryos, genes associated with abnormal mesoderm and dorsal–ventral axis formation, zygotic genome activation (ZGA), and anti-apoptosis were downregulated, whereas pro-apoptotic genes were upregulated, which may contribute to the higher abnormal mortality observed during embryonic development. Overall, this study demonstrates optimized conditions for inducing triploidy in grass carp via cold shock and provides insights into the transcriptomic changes that take place in cold shock-induced embryos, which could inform future grass carp genetic breeding programs. Full article

Polyploidy plays a significant role in loquat breeding, particularly in triploid breeding for seedless fruit production. Currently, loquat polyploid breeding primarily relies on natural seedling selection and sexual hybridization approaches. In this study, unfertilized ovules from four loquat varieties were in vitro cultured. Gynogenesis and embryoid regeneration were achieved in ‘Xingning 1’ and ‘Huabai 1’, with ‘Xingning 1’ demonstrating the highest gynogenesis efficiency (21.63%). Flow cytometry and chromosome counting revealed that the obtained embryoid lines included haploid, diploid, tetraploid, hexaploid, and chimeric ploidy types. Further characterization of ‘Xingning 1’-derived embryoid lines through SSR markers and whole-genome resequencing confirmed that the haploid, diploid, tetraploid, and hexaploidy embryoid originated from haploid–somatic chimeras, diploid, doubled diploid and tripled diploid, respectively. Metabolic analysis showed a positive correlation between ploidy level and the content of both soluble sugars and organic acids. This study explored a novel platform for polyploid induction in loquat and may provide methodological insights for improvement of other perennial fruit trees. Full article

The Chinese jujube (Ziziphus jujuba Mill.), an economically significant fruit tree native to China, is valued for both fresh and dried uses. In plants, 2n gametes serve as the fundamental basis for creating a sexual polyploid germplasm. This study investigated the 2n gametogenesis frequency in triploid hybrid jujubes through meiotic analysis of the hybrid strain Q161 and a two-year pollen analysis on hybrid progeny, assessing the natural 2n pollen frequencies to identify a high-2n-pollen germplasm and revealing the occurrence of 2n pollen. Meiotic analysis of the triploid hybrid Q161 (2n = 36) revealed cytological anomalies, including binucleate cells (22.80% abnormal tetrads), with natural 2n pollen production rates reaching 4.00% and 4.67% over two consecutive years. Scanning electron microscopy (SEM) revealed that the 2n pollen exhibited pronounced exine ornamentation with cerebroid sculpturing and tubercle-like structures at the apertures. Analysis of the triploid progeny for two consecutive years demonstrated a pollen viability of 30.45% and 23.83% (CV: 19. 39–29.69%), with the mean 2n pollen frequencies of 22.52% and 7.64%, peaking at 52.16% and 28.95% in elite individuals. Six triploid germplasm accessions with naturally elevated 2n pollen frequencies were identified. Under natural conditions, a triploid hybrid germplasm in Chinese jujube produces 2n pollen grains due to abnormal meiotic behavior, and a natural triploid germplasm with high pollen productivity was identified. This research provides a critical theoretical foundation for sexual polyploid breeding strategies. Full article

Lantana camara L. is sold worldwide for ornamental purposes, although it is also characterized by high invasiveness potential. Genetic and molecular data available for L. camara are still poor, and breeding is performed through conventional methods. This study focused on a molecular genotyping analysis through the ddRADseq method on an experimental collection of lantana clonal lines to evaluate the potential of molecular techniques in performing marker-assisted breeding, in favour of variety registration and in guaranteeing plant variety protection for the species. Although high genetic uniformity was observed in the population, a unique molecular profile was assigned to every line, indicating the effectiveness of the approach used. Interestingly, low degrees of heterozygosity were observed. In addition, the possibility of inferring ploidy levels through SNP profiles was assessed since it would avoid the necessity of previous biological knowledge and the use of fresh materials. Ploidy analysis is of high interest for lantana breeding to obtain less invasive triploids. Flow cytometry and chromosome counting were used for inference assessment. An nQuack framework provided correct results for the majority of the clonal lines, confirming its effectiveness. These findings encourage the adoption of molecular systems to help breed minor species such as L. camara. Full article

Crossing between selected lines could combine the additive genetic variance accumulated within the lines and the non-additive genetic variance between the lines in the genetic improvement of aquatic animals, thus obtaining progeny with favorable traits. However, the value of this breeding strategy has not been investigated in the Fujian oyster Crassostrea angulata due to the lack of available sufficiently selected lines. In this study, a complete 3 × 3 diallel cross was established between the normal (N), golden (G), and black (B) shell lines of C. angulata. The growth (shell height and living weight) and survival of three purebred groups and six hybrid groups under three environments (Luoyu, Jiangkou, and Houhai) were comprehensively evaluated during the larval and grow-out stages. The general combining ability (GCA) of the parental lines and the specific combining ability (SCA) of the hybrid groups were also estimated. The shell heights of the hybrid groups were significantly lower than those of their parental groups on day 25, exhibiting negative heterosis (MPH: −13.01 to −1.45; HPH: −16.69 to −5.76). Meanwhile, the survival rates of the hybrids were significantly higher than those of the parental groups. A negative value of SCA was recorded for NG (N♀ × G♂) (−0.031), which was in line with its lower survival rate on day 25. Significant heterosis was observed in the growth and survival of each hybrid group during the grow-out stage. The shell height and living weight of the hybrid groups were significantly higher in Houhai than in Luoyu and Jiangkou, but the survival rate in Houhai was significantly lower than in the other two sites. BG (B♀ × G♂) and GB (G♀ × B♂) had higher SCA values than the other four groups, which was consistent with their superior traits. The heterosis of shell height, living weight, and survival rate of BG was significantly greater than in the other five hybrid groups, which could be used as potential parents for breeding high-quality diploid and triploid Fujian oysters. This study demonstrated that the traits of C. angulata could be significantly improved by crossing between different selected lines, providing a reference for evaluating the utilization value of non-additive genetic effects (heterosis) between selected lines in bivalves. Full article

Encyclia Hook. is a tropical orchid known for its delightful fragrance and distinctive flower shapes, which have been widely used as superior parents for inter and intrageneric breeding. In this study, karyotype analysis shows that the number of chromosomes in diploid plants of Encyclia is 40, with all species exhibiting mesial and central filamentous chromosomes. The karyotype of diploid plants is primarily 2B. Among the three detected intergeneric hybrids, both Enanthleya Circus Lady ‘Coastal Star’ and Guaricyclia Kyoguchi ‘Fumi’ are triploid, while Robertsara ‘Green Wonder’ is tetraploid. By evaluating various lysates and tissues, a flow cytometry system (FCM) is developed with Galbraith’s buffer (GLB) and tender leaves. By using Dendrobium officinale as an external reference, the genome size of the Encyclia species and its intra and intergeneric hybrids are assessed, which ranges from 1.07 Gb to 3.23 Gb. These results will establish a foundation for high-throughput identification of germplasms, advance crossbreeding efforts, and enhance the understanding of the genome in Encyclia. Full article

Idesia polycarpa from Sichuan is a valuable germplasm with high economic potential, but it faces variety scarcity. To address this, this study collected 16 varieties (lines), identifying IpHT1 as a promising parent due to its high oil content (38.5%) and red fruits. Polyploid induction via adding 0.50% colchicine to Murashige and Skoog (MS) medium yielded 520 IpHT1 mutagenized seedlings. Subsequently, flow cytometry (FCM) was performed on 401 morphologically variant seedlings which had been initially screened, resulting in the identification of 15 suspected triploids, 35 suspected tetraploids, and 3 chimeras. Furthermore, fluorescence in situ hybridization (FISH) analysis found that the probe (AG₃T₃)₃ had terminal signals at both ends of each chromosome, allowing for the counting of 42 chromosomes in diploids and 84 in tetraploids. The probe 5S rDNA showed 2, 3, and 4 hybridization signals in the interphase nuclei of diploid, triploid, and tetraploid cells, respectively, but the probe (GAA)₆ failed to produce any signal on I. polycarpa chromosomes. Ultimately, 18 polyploids were selected, including 7 triploids and 11 tetraploids. Triploids and tetraploids showed significant leaf morphological and physiological differences from diploids. Consequently, this study successfully established a polyploid breeding system for I. polycarpa, thereby enhancing its genetic diversity and breeding potential. Full article

Banana breeding consists of obtaining diploid, triploid, and tetraploid intra- and interspecific hybrids by conventional breeding methods with the objective of aggregating characteristics of agronomic and commercial interest. Given the narrow genetic base of bananas, Embrapa’s Banana Genetic Breeding Program (BGBP) aims at crosses between improved diploids (ID) (ID × ID) and between improved diploids (ID) and commercial triploids (ID × CTP) and tetraploids (ID × CTT), in order to increase the genetic base and variability in bananas regarding agronomic traits of interest and resistance to main biotic and abiotic factors. These improved diploids are resistant to main fungal diseases such as yellow (YSD) and black Sigatoka (BSD) disease and Fusarium wilt (race 1 and subtropical race 4), the latter being one of the most devastating diseases in bananas. The genetic diversity between 22 improved diploids and seven commercial banana triploids was analyzed using DNA molecular markers. Five IRAP (Inter-Retrotransposon Amplified Polymorphism, 7 ISSR (Inter-Simple Sequence Repeats) and 12 SSR (Simple Sequence Repeat) markers were used. The genetic dissimilarity matrix was based on the Jaccard dissimilarity index; clusters were separated using the UPGMA (Unweighted Pair Group Method With Arithmetic Mean) method and cophenetic correlation of 0.8755. This study of the genetic diversity between improved diploids and commercial triploids, based on the genetic dissimilarity matrix, revealed that the most dissimilar diploids were DM23 and DM15 (74%) and DM16 and DM15 (74%). The smallest genetic distances between the improved diploids and commercial triploids were between TCGN25 and DM17 (50%) and TCN26 and DM17 (50%). The genetic distance matrix also revealed important genotypes to be used in crosses in order to maintain good characteristics in commercial triploids when crossed with improved diploids. The results of our study provide better breeding strategies for one of the largest banana-breeding programs worldwide focused on the development of banana varieties resistant to main biotic and abiotic factors. Full article

Leucaena is a versatile legume shrub/tree used as tropical livestock forage and in timber industries, but it is considered a high environmental weed risk due to its prolific seed production and broad environmental adaptation. Interspecific crossings between Leucaena species have been used to create non-flowering or sterile triploids that can display reduced weediness and other desirable traits for broad use in forest and agricultural settings. However, assessing the success of the hybridisation process before evaluating the sterility of putative hybrids in the target environment is advisable. Here, RNA sequencing was used to develop breeding markers for hybrid parental identification in Leucaena. RNA-seq was carried out on 20 diploid and one tetraploid Leucaena taxa, and transcriptome-wide unique genetic variants were identified relative to a L. trichandra draft genome. Over 16 million single-nucleotide polymorphisms (SNPs) and 0.8 million insertions and deletions (indels) were mapped. These sequence variations can differentiate all species of Leucaena from one another, and a core set of about 75,000 variants can be genetically mapped and transformed into genotyping arrays/chips for the conduction of population genetics, diversity assessment, and genome-wide association studies in Leucaena. For genetic fingerprinting, more than 1500 variants with even allele frequencies (0.4–0.6) among all species were filtered out for marker development and testing in planta. Notably, SNPs were preferable for future testing as they were more accurate and displayed higher transferability within the genus than indels. Hybridity testing of ca. 3300 putative progenies using SNP markers was also more reliable and highly consistent with the field observations. The developed markers pave the way for rapid, accurate, and cost-effective diversity assessments, variety identification and breeding selection in Leucaena. Full article

Near-infrared spectroscopy (NIR) has become an essential tool for non-destructive analysis in various fields, including aquaculture. This study presents a pioneering application of portable NIR spectrometers to analyze glycogen content in the gonadal tissues of the Pacific oyster (Crassostrea gigas), marking the first instance of developing quantitative models for glycogen in tetraploid C. gigas. The research also provides a comparative analysis with models for diploid and triploid oysters, underscoring the innovative use of portable NIR technology in aquaculture. Two portable NIR spectrometers were employed: the Micro NIR 1700 (908–1676 nm) and the Micro PHAZIR RX (1624–2460 nm). Near-infrared spectra were acquired from the gonadal tissues of diploid, triploid, and tetraploid C. gigas. Quantitative models for glycogen content were developed and validated using cross-validation methods. Additionally, qualitative models for different ploidies and genders were established. For the Micro NIR 1700, the cross-validation correlation coefficients (Rcv) and cross-validation relative predictive errors (RPDcv) for glycogen were 0.949 and 3.191 for diploids, 0.915 and 2.498 for triploids, and 0.902 and 2.310 for tetraploids. The Micro PHAZIR RX achieved Rcv and RPDcv values of 0.781 and 2.240 for diploids, 0.839 and 2.504 for triploids, and 0.717 and 1.851 for tetraploids. The Micro NIR 1700 demonstrated superior quantitative performance, with RPD values exceeding 2, indicating its effectiveness in predicting glycogen content across different ploidy levels. Qualitative models showed a performance index of 91.6 for diploid and 95 for tetraploid genders using the Micro NIR 1700, while the Micro PHAZIR RX achieved correct identification rates of 99.79% and 100% for diploid and tetraploid genders, respectively. However, differentiation of ploidies was less successful with both instruments. This study’s originality lies in establishing the first quantitative models for glycogen content in tetraploid C. gigas using portable NIR spectrometers, highlighting the significant advancements in non-destructive glycogen analysis. The applicability of these findings is substantial for oyster breeding programs focused on enhancing meat quality traits. These models provide a valuable phenotyping tool for selecting oysters with optimal glycogen content, demonstrating the practical utility of portable NIR technology in aquaculture. Full article

As one of the most influential environmental factors, drought stress greatly impacts the development and production of plants. Triploid-induced Passiflora edulis Sims ‘Mantianxing’ is an important new cultivar for multi-resistance variety selective breeding, which is one of the P. edulis breeding essential targets. However, the performance of triploid ‘Mantianxing’ under drought stress is unknown. In order to study the drought resistance of triploid ‘Mantianxing’, our study compared drought-related indicators in diploids and triploids under natural drought experiments, including morphological, physiological, and biochemical characteristics. Results showed that triploid P. edulis ‘Mantianxing’ showed variable responses to drought treatment. Compared with diploids, triploids showed higher photosynthesis and chlorophyll fluorescence, osmotic adjustment substances, and antioxidant enzyme activity under drought stress and faster chlorophyll biosynthesis and growth recovery after rewatering. Generally speaking, these results indicate that the drought resistance of triploid P. edulis is superior to diploid. This study provides scientific information for breeding stress tolerance variety of P. edulis ‘Mantianxing’ new cultivar. Full article

Tetraploid oysters are artificially produced oysters that do not exist in nature. The successful breeding of 100% triploid oysters resolved the difficulties of traditional drug-induced triploids, such as the presence of drug residues and a low triploid induction rate. However, little is known concerning the biochemical composition and nutrient contents of such tetraploids. Therefore, we investigated compositional differences among diploid, triploid, and tetraploid Crassostrea gigas as well as between males and females of diploids and tetraploids. The findings indicated that glycogen, EPA, ∑PUFA, and omega-3 contents were significantly higher in triploid oysters than in diploids or tetraploids; tetraploid oysters had a significantly higher protein content, C14:0, essential amino acid, and flavor-presenting amino acid contents than diploids or triploids. For both diploid and tetraploids, females had significantly higher levels of glutamate, methionine, and phenylalanine than males but lower levels of glycine and alanine. In addition, female oysters had significantly more EPA, DHA, omega-3, and total fatty acids, a result that may be due to the fact that gonadal development in male oysters requires more energy to sustain growth, consumes greater amounts of nutrients, and accumulates more proteins. With these results, important information is provided on the production of C. gigas, as well as on the basis and backing for the genetic breeding of oysters. Full article

This study layed the groundwork for the creation of haploid and triploid celestial goldfish, presenting essential data derived from preliminary investigations. The research delved into three main areas: the short-term preservation of celestial goldfish gametes, the interaction between oocytes and foreign sperm, and the effects of temperature on fertilized eggs concerning hatching rates and late-stage ocular development. Initially, the study explored the optimal semen dilution ratio for celestial goldfish under microscopic examination. And the hybridization of the largemouth bass and celestial goldfish was investigated: largemouth bass sperm was crossbred with celestial goldfish eggs, and it was found that their sperm could not stimulate the development of celestial eye eggs. At last, celestial goldfish fertilized eggs were stimulated at 4 °C and −20 °C, respectively, to observe their impact on the hatching rate and later celestial eye rate. The results revealed no significant differences in hatching rate and celestial eye rate between the cold stimulus groups and the control group, but numerically, the 4 °C cold stimulation reduced the celestial eye rate of celestial goldfish fertilized eggs. The research provided fundamental data for artificial breeding and hybridization experiments in celestial goldfish. Full article

The sweet oranges [C. sinensis (L.) Osbeck] produced in India are mainly consumed fresh as a table fruit and in the form of freshly extracted juice. Currently, a fraction of the fruit is processed into products like orange juice, concentrates, pulp, and other value-added products. Seedless varieties are preferred both by the consumers and by the processing industry; however, indigenously developed seedless sweet orange cultivars are not available. Citrus triploids are usually seedless due to their abnormal meiosis and embryo abortion. A research study was undertaken at CCRI Nagpur to develop triploid seedless plants of the local sweet orange cultivar Mosambi through endosperm culture in the shortest possible time by dovetailing various techniques. Various steps, viz., endosperm excision, standardization of media for callus initiation, somatic embryogenesis, shoot/root differentiation, mini-grafting, and validation of the triploid status through flow cytometry, chromosome counting, and other morphological parameters, were standardized, and seven confirmed triploid plants were finally produced. An evaluation of fruit quality parameters during the 2022 and 2023 fruiting seasons revealed that the fruits of the triploid sweet orange trees were commercially seedless. This is the first reported comprehensive study on the successful development of commercially seedless plants of the sweet orange cultivar Mosambi. The fruits of the triploid plants showed desirable fruit quality parameters in terms of their seed number (3 to 5.9/fruit), higher vitamin C content (34.4 to 42.66 mg), and lower limonene content (7.77 to 11.34 µg/mL). These triploid plants have the potential to gain recognition as a distinct variety. Full article

The East African Highland banana (Mutika/Lujugira subgroup) is composed of triploid (AAA) cooking and beer banana varieties that are adapted to the high-altitude region of the Great Lakes region of East Africa. Banana production is affected by several biotic and abiotic factors. Breeding opportunities in bananas are limited due to female sterility and parthenocarpy. The genetic diversity of crops enables breeders to develop new germplasm. Molecular markers have been used widely to dissect crop plants’ genetic diversity. This study assessed the genetic variation in 27 varieties from the Mutika/Lujugira subgroup using random amplified polymorphic DNA (RAPD). No genetic variation was observed among the banana varieties, and the 18 ten-mer primers produced monomorphic banding profiles. The genetic homogeneity of this banana subgroup is not congruent with their extensive morphological variation. Domestication and the bottleneck effect are often cited as the cause of reduced diversity in crop plants. On the other hand, several mechanisms, including somatic mutations, transposable elements, polyploidy, genome plasticity, and epigenetic mechanisms, are known to increase plant phenotypic variability. Further in-depth research is needed to explain the puzzle between the genetic and morphological diversity in the Mutika/Lujugira subgroup. Full article