Search Results (19)

Yak is an excellent germplasm resource on the Tibetan Plateau and is able to live in high-altitude areas with hypoxic, cold, and harsh environments. Studies on induced pluripotent stem cells (iPSCs) in large ruminants commonly involve a combination strategy involving six transcription factors, Oct4, Sox2, Klf4, c-Myc, Nanog, and Lin28 (OSKMNL). This strategy tends to utilize genes from the same species to optimize pluripotency maintenance. In this study, we cloned the six pluripotency genes (OSKMNL) from yak and constructed a multi-cistronic lentiviral vector carrying these genes. This vector efficiently delivered the genes into yak fibroblasts, aiming to promote the reprogramming process. We verified that the treated cells had several pluripotency characteristics, marking the first successful construction of a lentiviral system carrying yak pluripotency genes. This achievement lays the foundation for subsequent establishment of yak iPSCs and holds significant implications for yak-breed improvement and germplasm-resource conservation. Full article

Castanopsis hystrix, a precious tree species in Southeast Asia, has the advantages of rapid growth and high-quality wood materials. However, there are problems such as its long breeding cycle and low efficiency, and being time-consuming, which greatly restricts the industrial development of C. hystrix. Performing genome selection (GS) for growth and wood traits for the early selection of superior progeny has great significance for the rapid breeding of new superior varieties of C. hystrix. We used 226 clones in the main distribution and 479 progenies within 23 half-sib families as experimental materials in this study. Genotyping datasets were obtained by high-throughput re-sequencing technology, and GS studies were conducted on the growth (tree height (H), diameter at breast height (DBH)) and wood (wood density (WD), fiber length (FL), and fiber length–width ratio (LWR)) traits. The coefficient of variation (CV) of five phenotypic traits ranged from 10.1% to 22.73%, the average CV of growth traits was 19.93%, and the average CV of wood traits was 9.72%. The Pearson correlation coefficients between the five traits were almost all significantly positive. Based on the Genomic Best Linear Unbiased Prediction (GBLUP) model, the broad-sense heritabilities of growth traits were higher than those of wood quality traits, and the different number of SNPs had little effect on the heritability estimation. GS prediction accuracy first increased and then reached a plateau at around 3K SNPs for all five traits. The broad-sense heritability of these five traits was significantly positively correlated with their GS predictive ability (r = 0.564, p < 0.001). Bayes models had better GS prediction accuracy than the GBLUP model. The 15 excellent progeny individuals were selected, and their genetic gain ranged from 0.319% to 2.671%. These 15 superior offspring individuals were 4388, 4438, 4407, 4468, 4044, 4335, 4410, 4160, 4212, 4461, 4052, 4014, 4332, 4389, and 4007, mainly from three families F5, F6, and F11. Our research lays out the technical and material foundation for the rapid breeding of new superior varieties of C. hystrix in southern China. Full article

Lilium concolor var. pulchellum has a brilliant flower colour, high germination rate, and resistance to cold, drought, and salinity and is an excellent source of lily germplasm. Anthocyanins are important flavonoids commonly found in plants and can make the flowers and fruits of plants more colourful. We first found that 0.2 mg/L 1-naphthaleneacetic acid (NAA) specifically induced the accumulation of anthocyanins, which were mainly cyanidins, in callus tissue culture of Lilium. Transcriptomic results indicated that anthocyanin accumulation was mainly involved in the flavonoid pathway, and an LhMYB1 transcription factor encoding 267 amino acids positively associated with anthocyanin accumulation was cloned from the MYB family. Subcellular localisation in tobacco showed that the gene was located in the nucleus of epidermal cells. Virus-induced gene silencing showed that silencing of the LhMYB1 gene on lily petals resulted in a purple to white colour change and a decrease in anthocyanin deposition, mainly in the upper and lower epidermis of the petals. Therefore, the results of this study will provide some ideas for the molecular breeding of lily flower colour. Full article

Iris laevigata Fisch. is an excellent ornamental plant in cold regions due to its unique ornamental ability and strong cold resistance. However, the flowering period of the population is only about 20 days, greatly limiting its potential uses in landscaping and the cutting flower industry. In addition, I. laevigata is often challenged with various abiotic stresses including high salinity and drought in its native habitats. Thus, breeding novel cultivars with delayed flowering time and higher resistance to abiotic stress is of high importance. In this study, we utilized sequencing data from the I. laevigata transcriptome to identify WRKYs and characterized IlWRKY22, a key transcription factor that modulates flowering time and abiotic stress responses. IlWRKY22 is induced by salt and drought stress. We cloned IlWRKY22 and found that it is a Group IIe WRKY localized in the nucleus. Overexpressing IlWRKY22 in Arabidopsis thaliana (L.) Heynh. and Nicotiana tabacum L. resulted in a delayed flowering time in the transgenic plants. We created transgenic N. tabacum overexpressing IlWRKY22, which showed significantly improved resistance to both salt and drought compared to the control plants. Thus, our study revealed a unique dual function of IlWRKY22, an excellent candidate gene for breeding novel Iris cultivars of desirable traits. Full article

To improve the ecological and economic benefits of Korean pine (Pinus koraiensis), we analysed and evaluated its germplasm resources. This promotes in-depth research and utilisation of germplasm resources, providing excellent genetic resources for Korean pine breeding. We performed genetic analysis based on morphological and physiological traits and nuclear SSR molecular marker data was performed by collecting 314 clones (5 ramets of each clone) of Korean pine from eight (8) locations within the Korean pine range. The core collection underwent testing and evaluation for representativeness using variable rate (VR), coincidence rate (CR), variance difference percentage (VD), mean difference percentage (MD), Shannon index (I), and other indicators. The results indicated significant differences in morphological and physiological traits among the populations. All traits had a coefficient of variation (CV) greater than 10%, except for the water content of the needles (WC), which had an average CV of 17.636%. The populations showed high overall genetic diversity, with the HL (Helong) population exhibiting the highest genetic diversity, with an Ne (number of effective alleles), I, and He (expected heterozygosity) of 3.171, 1.103, and 0.528, respectively. Genetic variation mainly originated from individuals within populations, while the variation between populations was relatively small, at only 3%. The population did not exhibit any distinct subpopulation structures and was mainly derived from two admixed gene pools. Six core sets were obtained using different sampling strategies, and subset 6 was identified as the core collection, consisting of 114 individuals, representing a selection rate of 36.31%. In conclusion, the most appropriate method for constructing the core collection of Korean pines is the M-strategy (maximizing the number of alleles), based on both phenotypic and molecular data. The resulting core collection effectively represents the genetic diversity of the entire population effectively. Full article

Clubroot disease, caused by Plasmodiophora brassicae, poses a significant global threat to cruciferous crops. The epidemic area of clubroot disease is expanding rapidly. In response to this pressing issue, there is a compelling need for the development of clubroot disease-resistant radish cultivars. China boasts an extensive array of radish varieties and germplasm resources. However, a comprehensive assessment of their resistance to clubroot has not yet been carried out, thereby impeding the effective utilization of germplasm and clubroot-resistant breeding. Therefore, it is urgent to systematically evaluate the clubroot resistance of the radish germplasm and identify resistant resources. In this study, clubroot resistance evaluations were conducted on 268 excellent radish varieties derived from 30 provinces in China, as well as seven accessions from Russia, North Korea, France, South Korea, and Germany. The resistance evaluation revealed a diverse range of resistance indices, with a mean disease index (DI) ranging from 0.6 to 58.5, showing significant disparities in clubroot resistance among these radish resources. A total of six accessions were characterized as highly resistant to clubroot, and a further 50 accessions were characterized as resistant. The disease-resistant radishes showed diversity in horticultural traits. Provinces in South China contributed significantly more resistance germplasm than those of North China. These materials are of great value for both genetic investigation and the crop breeding of clubroot resistance. Furthermore, we employed a previously established clubroot-resistance-linked SSR marker to analyze the clubroot-resistant resources. The accessions exhibited dissimilar genetic profiles from known clubroot-resistant germplasm, suggesting their potential status as novel sources of clubroot resistance. Conclusively, these newly identified accessions enriched the genetic diversity within the clubroot-resistant gene pool and may contribute to the future cloning of previously undiscovered clubroot-resistant genes. Full article

Alnus cremastogyne is a rapidly growing broad-leaved tree species that is widely distributed in southwest China. It has a significant economic and ecological value. However, with the expansion of the planting area, the influence of phenotypic variation and differentiation on Alnus cremastogyne has increased, resulting in a continuous decline in its genetic quality. Therefore, it is crucial to investigate the phenotypic variation of Alnus cremastogyne and select excellent breeding materials for genetic improvement. Herein, four growth-related phenotypic traits (diameter at breast height, the height of trees, volume, height under the branches) and twelve reproductive-related phenotypic traits (fresh weight of single cone, dry weight of single cone, seed weight per plant, thousand kernel weight, cone length, cone width, cone length × cone width, fruit shape index, seed rate, germination rate, germination potential, germination index) of 40 clones from four provenances were measured and analyzed. The phenotypic variation was comprehensively evaluated by correlation analysis, principal component analysis and cluster analysis, and excellent clones were selected as breeding materials. The results revealed that there were abundant phenotypic traits variations among and within provenances. Most of the phenotypic traits were highly significant differences (p < 0.01) among provenances. The phenotypic variation among provenances (26.36%) was greater than that of within provenances clones (24.80%). The average phenotypic differentiation coefficient was accounted for 52.61% among provenances, indicating that the phenotypic variation mainly came from among provenances. The coefficient of variation ranged from 9.41% (fruit shape index) to 97.19% (seed weight per plant), and the repeatability ranged from 0.36 (volume) to 0.77 (cone width). Correlation analysis revealed a significantly positive correlation among most phenotypic traits. In principal component analysis, the cumulative contribution rate of the first three principal components was 79.18%, representing the main information on the measured phenotypic traits. The cluster analysis revealed four groups for the 40 clones. Group I and group II exhibited better performance phenotypic traits as compared with group III and group IV. In addition, the four groups are not clearly clustered following the distance from the provenance. Employing the multi-trait comprehensive evaluation method, 12 excellent clones were selected, and the average genetic gain for each phenotypic trait ranged from 4.78% (diameter at breast height) to 32.05% (dry weight of single cone). These selected excellent clones can serve as candidate materials for the improvement and transformation of Alnus cremastogyne seed orchards. In addition, this study can also provide a theoretical foundation for the genetic improvement, breeding, and clone selection of Alnus cremastogyne. Full article

As a “living fossil”, ginkgo (Ginkgo biloba L.) has significant ornamental, medicinal, and timber value. However, the breeding improvement of ginkgo was limited by the lack of enough excellent germplasms and suitable molecular markers. Here, we characterized numerous polymorphic insertion/deletion (InDel) markers using RAD-seq in 12 different ginkgo cultivars. The total of 279,534 InDels identified were unequally distributed across 12 chromosomes in the ginkgo genome. Of these, 52.56% (146,919) and 47.44% (132,615) were attributed to insertions and deletions, respectively. After random selection and validation, 26 pairs of polymorphic primers were used for molecular diversity analysis in 87 ginkgo cultivars and clones. The average values of observed heterozygosity and polymorphism information were 0.625 and 0.517, respectively. The results of population structure analyses were similar to those of neighbor-joining and principal component analyses, which divided all germplasms into two distinct groups. Moreover, 11 ginkgo core collections accounted for approximately 12.64% of the total ginkgo germplasms obtained, representing well the allelic diversity of all original germplasms. Therefore, these InDels can be used for germplasm management and genetic diversity analyses in ginkgo and the core collections will be used effectively for ginkgo genetic improvement. Full article

The quality of wheat primarily depends on its storage protein quality, especially in regards to gluten content and high-molecular-weight glutenin subunits (HMW-GS). The number of HMW-GS alleles is limited in bread wheat (Triticum aestivum L.), whereas it is abundant in wheat relatives. Therefore, HMW-GS alleles from wheat relatives could provide a potential for improving quality in wheat breeding. Thinopyrum elongatum (EE) is one of the relatives of wheat. The E genome is closely related to the ABD genome in wheat; therefore, Th. elongatum is often used as an excellent exogenous gene donor for wheat genetic improvement. In this study, the high-molecular glutenin subunit gene was cloned and sequenced from Th. elongatum. A specific molecular marker for identifying the Glu-1Ey subunit gene was developed and applied to detected wheat-Th. elongatum alien introgression lines. Quality analysis indicated that the substitution and addition lines containing Th. elongatum alleles significantly (p < 0.05) increased grain protein content by 3.76% to 5.11%, wet-gluten content by 6.55% to 8.73%, flour 8-MW by 0.25% to 6.35%, and bread volume value by 33.77 mL to 246.50 mL, in comparing it with Chinese Spring. The GMP content and lactic acid SRC showed significant positive correlations with flour processing quality and might be used as indicators for wheat quality. The results were expected to provide a novel route for improving processing quality in wheat quality breeding. Full article

Acacia melanoxylon (blackwood) is a valuable wood with excellent-quality heartwood extensively utilized worldwide. The main aim of this study was to confirm the horizontal and vertical variation and provide estimated values of genetic gains and clonal repeatabilities for improving the breeding program of A. melanoxylon. Six blackwood clones at 10 years old were analyzed in Heyuan and Baise cities in China. Stem trunk analysis was conducted for sample trees to explore the differences between heartwood and sapwood. The heartwood radius (HR), heartwood area (HA), and heartwood volume (HV) in heartwood properties decreased as the tree height (H) in growth traits increased, and the HV = 1.2502 DBH (diameter at breast height)^1.7009 model can accurately estimate the heartwood volume. Furthermore, G × E analysis showed that the heritabilities of the eleven indices, including DBH, DGH (diameter at ground height), H, HR, SW (sapwood width), BT (bark thickness), HA, SA (sapwood area), HV, HRP (heartwood radius percentage), HAP (heartwood area percentage), and HVP (heartwood volume percentage) were between 0.94 and 0.99, and repeatabilities of the eleven indices were between 0.74 and 0.90. Clonal repeatability of DBH (0.88), DGH (0.88), and H (0.90) in growth traits and HR (0.90), HVP (0.90), and HV (0.88) in heartwood properties were slightly higher than for SA (0.74), SW (0.75), HAP (0.75), HRP (0.75), and HVP (0.75). These data also implied that the growth characteristics of heartwood and sapwood of blackwood clones were less affected by the environment and had substantial heritability. Full article

Psathyrostachys huashanica Keng (2n = 2x = 14, NsNs) is an excellent gene resource for wheat breeding, which is characterized by early maturity, low plant height, and disease resistance. The wheat-P. huashanica derivatives were created by the elite genes of P. huashanica and permeate into common wheat through hybridization. Among them, a long-glume material 20JH1155 was identified, with larger grains and longer spike than its parents. In the present study, the methods of cytological observation, GISH, and sequential FISH analysis showed that 20JH1155 contained 21 pairs of wheat chromosomes and a pair of P. huashanica. There were some differences in 5A and 7B chromosomes between 20JH1155 and parental wheat 7182. Molecular marker, FISH, and sequence cloning indicated 20JH1155 alien chromosomes were 3Ns of P. huashanica. In addition, differentially expressed genes during immature spikelet development of 20JH1155 and 7182 and predicted transcription factors were obtained by transcriptome sequencing. Moreover, a total of 7 makers derived from Ph#3Ns were developed from transcriptome data. Taken together, the wheat-P. huashanica derived line 20JH1155 provides a new horizon on distant hybridization of wheat and accelerates the utilization of genes of P. huashanica. Full article

Coffea canephora has high but inadequately exploited genetic diversity. This diversity, if well exploited, can sustain coffee productivity amidst climate change effects. Drought and heat stress are major global threats to coffee productivity, quality, and tradable volumes. It is not well understood if there is a selectable variation for drought stress tolerance in Robusta coffee half-sibs as a result of watering deficit pre-exposure at the germination stage. Half-sib seeds from selected commercial clones (KR5, KR6, KR7) and a pipeline clone X1 were primed with deficit watering at two growth stages followed by recovery and later evaluated for tolerance to watering deficit stress in three different temperature environments by estimation of plant growth and wilt parameters. Overall, the KR7 family performed the best in terms of the number of individuals excelling for tolerance to deficit watering. In order of decreasing tolerance, the 10 most promising individuals for drought and heat tolerance were identified as: 14.KR7.2, 25.X1.1, 35.KR5.5, 36.KR5.6, 41.KR7.5, 46.KR6.4, 47.KR6.5, 291.X1.3, 318.X1.3, and 15.KR7.3. This is the first prospect into the potential of C. canephora half-sibs’ diversity as an unbound source of genetic variation for abiotic stress tolerance breeding. Full article

To select excellent clones characterized by fast growth and high-quality wood of Catalpa fargesii Bur., growth traits, including height and diameter at breast height (DBH) at 1, 9, 11 and 13 years old, were collected and wood properties, including the Pilodyn value and elastic modulus at age 13 of 200 clones of C. fargesii from different ecological areas (four provinces), were measured. Genetic variation analysis and repeatability estimation were carried out. The correlation between the characteristics and the correlation between the characteristics and the geographical and climatic factors were analyzed. Excellent clones were selected by a comprehensive evaluation method combined with breeding values. The traits of C. fargesii were significantly different among the clones. The coefficient of variation (CV) ranged from 12.12% to 26.17%, and the repeatability (R) ranged from 0.79 to 0.97. The growth traits among ages of 9, 11 and 13 were significantly correlated, and the growth traits at ages of 9, 11 and 13 and Pilodyn values at the age of 13 were positively correlated with altitude and negatively correlated with longitude. Tree growth at ages of 11 and 13 showed a high negative correlation with wood properties. The suitable tree age for the early selection of clones with excellent growth and wood properties is 11 years old. Nine clones with excellent growth, six clones with high-quality wood and three clones with great growth and wood properties were selected, which laid a foundation for further optimization of C. fargesii clones. Additionally, C. fargesii clones in the western and high-altitude areas featured fast growth, while those in the eastern and low-altitude areas had better wood properties. This study serves as a reference for the selection of C. fargesii clones in different ecological areas. Full article

The low temperature (LT) conditions that prevail during winter in subtropical regions of India drastically affect the growth and yield of sugarcane. To identify low-temperature-tolerant agronomical acceptable genotypes for immediate deployment as donor parents in the subtropical sugarcane breeding program, 34 sugarcane clones belonging to 7 genetically diverse groups were evaluated under three crop environments, viz., spring planting, winter ratoon and spring ratoon, during 2015–2016 and 2016–2017. In the winter ratoon crop, commercial cane sugar and cane yield were reduced, whereas sucrose % was increased over the spring planted crop and the spring ratoon crop. The wild species and introgressed hybrid groups showed improvement for yield and quality traits in the winter ratoon crop, whereas commercial and near commercial groups showed reduction for these traits over the plant and spring ratoon crops. The tropical cultivars group was the poorest performer irrespective of the traits and crops. Yield per se under a stress environment was adjudged as the best selection criteria. For classification of sugarcane clones according to their low temperature tolerance, an index named winter tolerance index (WTI) is proposed which takes into account the winter sprouting index (WSI), winter growth and yield per se of the winter ratoon crop. The WTI had significant positive association with WSI, cane yield, millable cane population and cane length. As per the WTI ratings, the wild species of Saccharum complex and introgressed hybrid groups were rated as excellent WT clones. Subtropical commercial or advanced generation groups were poor WT clones, and tropical commercial cultivars group were winter sensitive clones. Clones such as AS04-635, AS04-1687, IK76-48, GU07-2276, IND00-1040, IND00-1038 and IND00-1039 had excellent tolerance, and GU07-3849, AS04-245, Co 0238, AS04-2097 and GU07-3774 had good WTI scores. The variety, Co 0238, may be continued for cultivation under LT regions with prophylactic measurers for red rot, while other clones listed above may be utilized in subtropical breeding programs. Full article

Erianthus arundinaceus is a valuable gene reservoir for sugarcane improvement. However, insufficient molecular markers for high-accuracy identification and tracking of the introgression status of E. arundinaceus chromatin impede sugarcane breeding. Fortunately, suppression subtractive hybridization (SSH) technology provides an excellent opportunity for the development of high-throughput E. arundinaceus-specific molecular markers at a reasonable cost. In this study, we constructed a SSH library of E. arundinaceus. In total, 288 clones of E. arundinaceus-specific repetitive sequences were screened out and their distribution patterns on chromosomes were characterized by fluorescence in situ hybridization (FISH). A subtelomeric repetitive sequence Ea086 and a diffusive repetitive sequence Ea009, plus 45S rDNA-bearing E. arundinaceus chromosome repetitive sequence EaITS were developed as E. arundinaceus-specific molecular markers, namely, Ea086-128, Ea009-257, and EaITS-278, covering all the E. arundinaceus chromosomes for high-accuracy identification of putative progeny. Both Ea086-128 and Ea009-257 were successfully applied to identify the authenticity of F₁, BC₁, BC₂, BC₃, and BC₄ progeny between sugarcane and E. arundinaceus. In addition, EaITS-278 was a 45S rDNA-bearing E. arundinaceus chromosome-specific molecular marker for rapid tracking of the inherited status of this chromosome in a sugarcane background. Three BC₃ progeny had apparently lost the 45S rDNA-bearing E. arundinaceus chromosome. We reported herein a highly effective and reliable SSH-based technology for discovery of high-throughput E. arundinaceus-specific sequences bearing high potential as molecular markers. Given its reliability and savings in time and efforts, the method is also suitable for development of species-specific molecular markers for other important wild relatives to accelerate introgression of wild relatives into sugarcane. Full article