Search Results (58)

Maize is a vital global crop, serving as a fundamental component of global food security. To support sustainable maize production, the accurate classification of maize seeds—particularly distinguishing haploid from diploid types—is essential for enhancing breeding efficiency. Conventional methods relying on manual inspection or simple machine learning are prone to errors and unsuitable for large-scale data. To overcome these limitations, we propose Slot-Maize, a novel deep learning architecture that integrates Convolutional Neural Networks (CNN), Slot Attention, Gated Recurrent Units (GRU), and Long Short-Term Memory (LSTM) layers. The Slot-Maize model was evaluated using two datasets: the Maize Seed Dataset and the Maize Variety Dataset. The Slot Attention module improves feature representation by focusing on object-centric regions within seed images. The GRU captures short-term sequential patterns in extracted features, while the LSTM models long-range dependencies, enhancing temporal understanding. Furthermore, Grad-CAM was utilized as an explainable AI technique to enhance the interpretability of the model’s decisions. The model demonstrated an accuracy of 96.97% on the Maize Seed Dataset and 92.30% on the Maize Variety Dataset, outperforming existing methods in both cases. These results demonstrate the model’s robustness, generalizability, and potential to accelerate automated maize breeding workflows. In conclusion, the Slot-Maize model provides a robust and interpretable solution for automated maize seed classification, representing a significant advancement in agricultural technology. By combining accuracy with explainability, Slot-Maize provides a reliable tool for precision agriculture. Full article

As one of the most important staple crops in the world, rice plays a pivotal role in world food security. The creation of doubled haploids based on anther culture is an important technology for rice breeding. However, at present, rice anther culture technology still faces many problems, such as genotype dependency, especially genotypes of indica rice. In this study, fifteen rice genotypes, including twelve japonica rice genotypes and three indica rice genotypes, were randomly selected and used to study anther culture by using a modified M8 medium. The results showed that the total callus induction rates of these different rice genotypes ranged from 0.81 to 13.95%, with an average of 6.64%, while the callus induction rates calculated for the top ten highest callus inductions for each rice genotype ranged from 2.75 to 17.00%, with an average of 10.56%. There were varying gaps between the total callus induction rates and the callus induction rates in these different rice genotypes. The fact that the gaps for some rice genotypes were relatively large indicated that standard tiller or anther collection was not applicable to all rice genotypes and that there was still a lot of room for improvement in the callus induction rate of some rice genotypes through optimization of the sampling method. The plantlet regeneration rates ranged from 12.55 to 456.54%, with an average of 200.10%. Although there were many albinos from anther culture for some rice genotypes, these would still meet the requirement if the rice genotypes had higher callus induction rates or regeneration rates. The percentages of seed setting of regenerated green seedlings ranged from 14% to 84%, with an average of 48.73%. Genetic diversity analysis showed that the genetic background of these different rice genotypes was representative, and the phylogenetic tree and Principal Component Analysis (PCA) divided them into indica and japonica types. Therefore, in this study, an anther culture method suitable for both indica and japonica rice genotypes was established, which could improve doubled haploid breeding in rice. Full article

Cannabis (Cannabis sativa) is a versatile crop belonging to the Cannabaceae family, and is dioecious, typically with separate male and female plants. The flowers of female plants, especially the trichomes, accumulate relatively higher contents of cannabinoids compared with those of male plants. For this reason, to obtain seeds that are genetically female, it is desirable to induce the development of male flowers on a female plant that produces genetically female haploid gametes. Silver thiosulfate (STS) is a highly effective chemical for male flower induction. We investigated male flower induction in three commercial cultivars of female cannabis (Spectrum303, SuperwomanS1, and CBGambit) regarding the treatment frequency, stage of application, and concentration of STS applied as a foliar spray. All three cultivars showed adequate induction of male flowers in response to 1.5 mM STS applied at the early reproductive stage. In particular, SuperwomanS1 was most highly responsive to induction of male flowers, even when treated with 0.3 mM STS at the early reproductive stage. Treatment with three applications of STS was more effective compared with a single application, but a single application of 1.5 mM STS at the early reproductive stage was sufficient for male flower induction. A single STS application during the middle stage of reproductive growth was inadequate for induction of male flowers. However, 6 weeks after three applications of STS, CBGambit exhibited approximately 54% male flower induction at 0.3 mM STS, Spectrum303 showed approximately 56% induction at 3 mM STS, and SuperwomanS1 yielded approximately 26% induction at 1.5 mM (expressed as percentage of total number of individuals with the induced male flowers). Pollen stainability tests using KI-I₂ solution and Alexander’s staining showed high pollen viability with over 65% at different single STS concentrations, indicating that pollen grains induced by STS have sufficient viability for the self-pollination. This study demonstrated that different cultivars of cannabis respond diversely to different STS concentrations and highlighted the potential benefits of three STS applications during the middle reproductive stage for cannabis breeding. Full article

Multiple factors can affect the R1-nj purple kernel expression and seed set, reducing its efficiency in identifying haploids in maize. The complex interaction among the haploid inducer (HI), source germplasm (SG), and season (S) is inevitable in in vivo maize haploid induction but could be used through compatibility and stability tests. We tested five HI genotypes on 25 distinct source germplasm in two different seasons of tropical savanna in Thailand. The dry season was more suitable than the rainy season for haploid induction. We noticed varying degrees of R1-nj inhibition among the 25 tropical source germplasm, with some of them exhibiting significant issues with the R1-nj purple kernel expression. Therefore, using the R1-nj alone may not provide accurate ploidy identification in maize. Despite the intense R1-nj expression, haploid inducer BHI306 showed poor stability and compatibility with tropical source germplasm for pollination rate and seed set during the rainy season. The GGE biplot suggested KHI42 and KHI64 as the most compatible haploid inducers under their respective two different mega-source germplasm for the pollination rate and R1-nj seed set. These findings can guide breeders in selecting the most compatible and stable haploid inducers under varying conditions. Full article

The cytoplasm of Aegilops kotschyi is known for the induction of male sterility and haploidy in wheat. Both systems originally appeared rather simple, but manipulation of the standard chromosome constitution of the nuclear genome revealed additional interactions. This study shows that while there is little or no allelic variation at the main fertility restorer locus Rf^multi on chromosome arm 1BS, additional genes may also be involved in the nuclear–mitochondrial genome interactions, affecting not only male fertility but also the growth rate, from pollen competition for fertilization and early endosperm divisions all the way to seed size and plant maturity. Some of these effects appear to be of a sporophytic nature; others are gametophytic. Induction of parthenogenesis by a rye inducer in conjunction with the Ae. kotschyi cytoplasm is well known. However, here we show that the cytoplasmic-nuclear interactions affect all aspects of double fertilization: producing maternal haploids from unfertilized eggs, diploids from fertilized eggs or synergids, embryo-less kernels, and fertilized eggs without fertilization of the double nucleus in the embryo sack. It is unclear how frequent the inducers of parthenogenesis are, as variation, if any, is obscured by suppressors present in the wheat genome. Genetic dissection of a single wheat accession revealed five distinct loci affecting the rate of maternal haploid production: four acting as suppressors and one as an enhancer. Only when the suppressing haplotypes are confirmed may it be possible to the identify genetic variation of haploidy inducers, map their position(s), and determine their nature and the mode of action. Full article

Haploid breeding can shorten the breeding period of new maize varieties and is an important means to increase maize yield. In the breeding program, a large number of haploid seeds need to be screened, and this step is mainly achieved manually, which hinders the industrialization of haploid maize breeding. This article aims to develop a multispectral camera to identify the haploid seeds automatically. The camera was manufactured by replacing narrow-band filters of the ordinary CCD camera, and the RGB, 405 nm, 980 nm and 1050 nm images of haploid or diploid seeds were simultaneously captured (the characteristic wavelengths were determined according to color and high-oil markers of maize). The performance was tested using four maize varieties with the two genetic markers. The results show that the developed multispectral camera significantly improved the recognition accuracy of haploid maize seeds to 92.33%, 97.33%, 97% and 93.33% for the TYD1903, TYD1904, TYD1907 and TYD1908 varieties, respectively. The cameras in the near-infrared region (wavelengths of 980 nm and 1050 nm) achieved better performance for the varieties of high-oil marker, with an increase of 0.84% and 1.5%, respectively. These results demonstrate the strong potential of the multispectral imaging technology in the haploid seed identification of maize. Full article

Phenotypic plasticity is the ability of an individual genotype to express phenotype variably in different environments. This study investigated the plasticity of yield-related traits of bread wheat by utilising 225 doubled haploid (DH) lines developed from cv. Westonia and cv. Kauz, through two field trials in Western Australia. Plasticity was quantified via two previously published methods: responsiveness to varying ecological conditions and slopes of reaction norms. The spikelets/spike was the most plastic trait, with an overall plasticity of 1.62. The least plastic trait was grain protein content, with an overall plasticity of 0.79. The trait hierarchy based on phenotypic plasticity was spikelets/spike > thousand kernel weight > seed number > seed length > grain yield > grain protein content. An increase in yield plasticity of 0.1 was associated with an increase in maximum yield of 4.45 kg ha⁻¹. The plasticity of seed number and grain protein content were significantly associated with yield plasticity. The maximal yield was positively associated with spikelets/spike and grain yield, whereas it negatively associated with grain protein content. In contrast, the minimal yield was found to be negatively related to the plasticity of spikelets/spike and the plasticity of grain yield, whereas it was not related to grain protein content plasticity. Seed number and seed length exhibited plastic responses at the higher fertilisation state while remaining relatively stable at the lower fertilisation state for the wheat DH population. The finding of the current study will play a key role in wheat improvement under the changing climate. Seed length and seed number should be the breeding target for achieving stable yield in adverse environmental conditions. Full article

Banana breeding faces numerous challenges, such as sterility and low seed viability. Enhancing our understanding of banana genetics, notably through next-generation sequencing, can help mitigate these challenges. The genotyping datasets currently available from genebanks were used to decipher cultivated bananas’ genetic makeup of natural cultivars using genome ancestry mosaic painting. This article presents the application of this method to breeding materials by analyzing the chromosome segregation at the origin of ‘Gold Finger’ (FHIA-01), a successful improved tetraploid variety that was developed in the 1980s. First, the method enabled us to clarify the variety’s intricate genetic composition from ancestral wild species. Second, it enabled us to infer the parental gametes responsible for the formation of this hybrid. It thus revealed 16 recombinations in the haploid male gamete and 10 in the unreduced triploid female gamete. Finally, we could deduce the meiotic mechanism lying behind the transmission of unreduced gametes (i.e., FDR). While we show that the method is a powerful tool for the visualization and inference of gametic contribution in hybrids, we also discuss its advantages and limitations to advance our comprehension of banana genetics in a breeding context. Full article

Sugar beet is susceptible to Beet curly top virus (BCTV), which significantly reduces yield and sugar production in the semi-arid growing regions worldwide. Sources of genetic resistance to BCTV is limited and control depends upon insecticide seed treatments with neonicotinoids. Through double haploid production and genetic selection, BCTV resistant breeding lines have been developed. Using BCTV resistant (R) [KDH13; Line 13 and KDH4-9; Line 4] and susceptible (S) [KDH19-17; Line 19] lines, beet leafhopper mediated natural infection, mRNA/sRNA sequencing, and metabolite analyses, potential mechanisms of resistance against the virus and vector were identified. At early infection stages (2- and 6-days post inoculation), examples of differentially expressed genes highly up-regulated in the ‘R’ lines (vs. ‘S’) included EL10Ac5g10437 (inhibitor of trypsin and hageman factor), EL10Ac6g14635 (jasmonate-induced protein), EL10Ac3g06016 (ribosome related), EL10Ac2g02812 (probable prolyl 4-hydroxylase 10), etc. Pathway enrichment analysis showed differentially expressed genes were predominantly involved with peroxisome, amino acids metabolism, fatty acid degradation, amino/nucleotide sugar metabolism, etc. Metabolite analysis revealed significantly higher amounts of specific isoflavonoid O-glycosides, flavonoid 8-C glycosides, triterpenoid, and iridoid-O-glycosides in the leaves of the ‘R’ lines (vs. ‘S’). These data suggest that a combination of transcriptional regulation and production of putative antiviral metabolites might contribute to BCTV resistance. In addition, genome divergence among BCTV strains differentially affects the production of small non-coding RNAs (sncRNAs) and small peptides which may potentially affect pathogenicity and disease symptom development. Full article

The doubled haploid (DH) technology is employed worldwide in various crop-breeding programs, especially maize. Still, restoring tassel fertility is measured as one of the major restrictive factors in producing DH lines. Colchicine, nitrous oxide, oryzalin, and amiprophosmethyl are common chromosome-doubling agents that aid in developing viable diploids (2n) from sterile haploids (n). Although colchicine is the most widely used polyploidy-inducing agent, it is highly toxic to mammals and plants. Therefore, there is a dire need to explore natural, non-toxic, or low-toxic cheaper and accessible substitutes with a higher survival and fertility rate. To the best of our knowledge, the advanced usage of human anticancer drugs “Paclitaxel (PTX)” and “Caffeine–Taurine (CAF–T)” for in vivo maize haploids doubling is being disclosed for the first time. These two antimitotic and antimicrotubular agents (PTX and CAF–T) were assessed under various treatment conditions compared to colchicine. As a result, the maximum actual doubling rates (ADR) for PTX versus colchicine in maize haploid seedlings were 42.1% (400 M, 16 h treatment) versus 31.9% (0.5 mM, 24 h treatment), respectively. In addition, the ADR in maize haploid seeds were CAF–T 20.0% (caffeine 2 g/L + taurine 12 g/L, 16 h), PTX 19.9% (100 μM, 24 h treatment), and colchicine 26.0% (2.0 mM, 8 h treatment). Moreover, the morphological and physiological by-effects in haploid plants by PTX were significantly lower than colchicine. Hence, PTX and CAF–T are better alternatives than the widely used traditional colchicine to improve chromosome-doubling in maize crop. Full article

In cereal breeding, in vitro androgenesis methods are frequently applied to achieve doubled haploid (DH) plants. The aim of this study was to determine the effects of genotype (three registered varieties and eight F₁ crossing combinations) and induction medium (W14mf and P4mf) on anther cultures (ACs) of triticale (×Triticosecale Wittmack). Androgenesis was induced in the treatment of each tested genotype, and the genotype significantly influenced the efficiency of AC, including in embryo-like structures (ELSs), albinos, green plantlets, and transplanted plantlets. The utilized medium also had a significant effect on the number of ELSs, albinos, and transplanted plantlets. Both media were suitable for AC in triticale DH plant production. The efficiency of AC was higher when using the P4mf medium (103.7 ELS/100 anthers, 19.7 green plantlets/100 anthers) than when using the W14mf medium (90.0 ELS/100 anthers, 17.0 green plantlets/100 anthers). However, the green plantlet regeneration efficiency of microspore-derived structures was 18.0% when using the W14mf medium, while this value was 15.9% in the case of ELSs induced with the P4mf medium. After nursery seed evaluation and propagation (DH₁), the genetic homogeneity of the offspring generation (DH₂) was tested using a molecular genetic method. Most of the tested DH lines showed homogeneity and were progressed into a breeding program after agronomic selection. Some DH lines showed inhomogeneity, which could be explained by the outcross inclination of triticale. We would like to call breeders’ attention to the outcross character of triticale and emphasize the vigilant propagation and maintenance of the triticale DH lines in breeding programs. Due to the outcross nature of triticale, even in self-pollinated genotypes, breeders should focus on careful maintenance, along with isolation in the case of line propagations, in triticale breeding programs. Full article

Crop breeding is an important global strategy to meet sustainable food demand. CRISPR/Cas is a most promising gene-editing technology for rapid and precise generation of novel germplasm and promoting the development of a series of new breeding techniques, which will certainly lead to the transformation of agricultural innovation. In this review, we summarize recent advances of CRISPR/Cas technology in gene function analyses and the generation of new germplasms with increased yield, improved product quality, and enhanced resistance to biotic and abiotic stress. We highlight their applications and breakthroughs in agriculture, including crop de novo domestication, decoupling the gene pleiotropy tradeoff, crop hybrid seed conventional production, hybrid rice asexual reproduction, and double haploid breeding; the continuous development and application of these technologies will undoubtedly usher in a new era for crop breeding. Moreover, the challenges and development of CRISPR/Cas technology in crops are also discussed. Full article

Doubled haploid (DH) technology has become integral to maize breeding programs to expedite inbred line development and increase the efficiency of breeding operations. Unlike many other plant species that use in vitro methods, DH production in maize uses a relatively simple and efficient in vivo haploid induction method. However, it takes two complete crop cycles for DH line generation, one for haploid induction and the other one for chromosome doubling and seed production. Rescuing in vivo induced haploid embryos has the potential to reduce the time for DH line development and improve the efficiency of DH line production. However, the identification of a few haploid embryos (~10%) resulting from an induction cross from the rest of the diploid embryos is a challenge. In this study, we demonstrated that an anthocyanin marker, namely R1-nj, which is integrated into most haploid inducers, can aid in distinguishing haploid and diploid embryos. Further, we tested conditions that enhance R1-nj anthocyanin marker expression in embryos and found that light and sucrose enhance anthocyanin expression, while phosphorous deprivation in the media had no affect. Validating the use of the R1-nj marker for haploid and diploid embryo identification using a gold standard classification based on visual differences among haploids and diploids for characteristics such as seedling vigor, erectness of leaves, tassel fertility, etc., indicated that the R1-nj marker could lead to significantly high false positives, necessitating the use of additional markers for increased accuracy and reliability of haploid embryo identification. Full article

A crucial aim for rapeseed breeders is increasing the seed yield through improving yield- and silique-related traits. In this research, traits including the number of seeds per silique, silique length, silique width, and thousand-seed weight combined with seed coat color were investigated. Yellow-seeded lines of oilseed rape have a thinner seed coat, lower fiber content, and higher protein and oil content than traditional varieties. Hybrids of yellow- and black-seeded Brassica napus, which combine the advantages of both types of seeds, are tested for their suitability in breeding. Therefore, the aim of this study was to assess a population of 78 doubled haploid lines with different seed coat colors to select genotypes connecting a light color of seeds with good yielding characteristics. Multivariate methods such as analysis of variance, correlation analysis, regression analysis, canonical variate analysis, Mahalanobis distances, and estimation of gene effects were used. The results showed that all tested genotypes were significantly different regarding all five studied traits. The multidimensional analyses gave an accurate overview of the relationship between the examined features. The color and weight of seeds were considered the most important for breeding rapeseed with better seed properties. Although a negative correlation between these two characteristics occurs quite commonly, it was not present in our plant material, which allowed us to select lines with a-light seed color and high thousand-seed weight, which is a significant achievement. As a result, two doubled haploid lines were chosen as valuable breeding material for creating oilseed rape varieties with improved biochemical characteristics of seeds. Full article

Anther culture is an efficient biotechnological tool in modern plant breeding programs to produce new varieties and parental lines in hybrid seed productions. However, some bottlenecks—low induction rate, genotype dependency, albinism—restrict the widespread utilization of in vitro anther culture in rice breeding, especially in Oryza sativa ssp. indica (indica) genotypes, while an improved efficient protocol can shorten the process of breeding. Three different induction media (N₆NDK_, N₆NDZ, Ali-1) and four plant regeneration media (mMSNBK1, MSNBK3, MSNBKZ1, MSNBKZ2) were tested with five indica rice genotypes to increase the efficiency of in vitro androgenesis (number of calli and regenerated green plantlets). The production of calli was more efficient on the N₆NDK medium with an average 88.26 calli/100 anthers and N₆NDZ medium with an average of 103.88 calli/100 anthers as compared to Ali-1 with an average of 6.96 calli/100 anthers. The production of green plantlets was greater when calli was produced on N₆NDK medium (2.15 green plantlets/100 anthers) compared when produced on to N₆NDZ medium (1.18 green plantlets/100 anthers). Highest green plantlets production (4.7 green plantlets/100 anthers) was achieved when mMSNBK1 plant regeneration medium was used on calli produced utilizing N₆NDK induction medium. In the best overall treatment (N₆NDK induction medium and mMSNBK1 plant regeneration medium), four tested genotypes produced green plantlets. However, the genotype influenced the efficiency, and the green plantlets production ranged from 0.4 green plantlets/100 anthers to 8.4 green plantlets/100 anthers. The ploidy level of 106 acclimatized indica rice plantlets were characterized with flow cytometric analyses to calculate the percentage of spontaneous chromosome doubling. Altogether, 48 haploid-, 55 diploid-, 2 tetraploid- and 1 mixoploid plantlets were identified among the regenerant plantlets, and the spontaneous chromosome doubling percentage was 51.89%. Utilization of DH plants have been integrated as a routine method in the Hungarian rice breeding program. The tetraploid lines can be explored for their potential to offer new scopes for rice research and breeding directions in the future. Full article