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[学术文献 ] CRISPR/Cas9-mediated editing of double loci of BnFAD2 increased the seed oleic acid content of rapeseed (Brassica napus L.) 进入全文
Front Plant Sci
Seed oleic acid is an important quality trait sought in rapeseed breeding programs. Many methods exist to increase seed oleic acid content, such as the CRISPR/Cas9-mediated genome editing system, yet there is no report on seed oleic acid content improvement via this system’s precise editing of the double loci of BnFAD2. Here, a precise CRISPR/Cas9-mediated genome editing of the encoded double loci (A5 and C5) of BnFAD2 was established. The results demonstrated high efficiency of regeneration and transformation, with the rapeseed genotype screened in ratios of 20.18% and 85.46%, respectively. The total editing efficiency was 64.35%, whereas the single locus- and double locus-edited ratios were 21.58% and 78.42%, respectively. The relative proportion of oleic acid with other fatty acids in seed oil of mutants was significantly higher for those that underwent the editing on A5 copy than that on C5 copy, but it was still less than 80%. For double locus-edited mutants, their relative proportion of oleic acid was more than 85% in the T1 and T4 generations. A comparison of the sequences between the double locus-edited mutants and reference showed that no transgenic border sequences were detected from the transformed vector. Analysis of the BnFAD2 sequence on A5 and C5 at the mutated locus of double loci mutants uncovered evidence for base deletion and insertion, and combination. Further, no editing issue of FAD2 on the copy of A1 was detected on the three targeted editing regions. Seed yield, yield component, oil content, and relative proportion of oleic acid between one selected double loci-edited mutant and wild type were also compared. These results showed that although the number of siliques per plant of the wild type was significantly higher than those of the mutant, the differences in seed yield and oil content were not significant between them, albeit with the mutant having a markedly higher relative proportion of oleic acid. Altogether, our results confirmed that the established CRISPR/Cas9-mediated genome editing of double loci (A5 and C5) of the BnFAD2 can precisely edit the targeted genes, thereby enhancing the seed oleic acid content to a far greater extent than can a single locus-editing system.
[学术文献 ] Identification of a Leafy Head Formation Related Gene in Chinese Cabbage (Brassica rapa L. ssp. pekinensis) 进入全文
Horticulturae
Leafy head formation is one of the most important characteristics of Chinese cabbage, and the process is regulated by a series of genes and environmental factors. In this study, a non-heading short leaf mutant slm was identified from an ethyl methane sulfonate mutagenesis (EMS) population of the heading Chinese cabbage line FT. The most significant phenotypic characteristics of slm was shortening leaves and increasing leaf numbers, which led to failure to form a leafy head. Genetic analysis showed that a single recessive gene Brslm was responsible for the mutant phenotype. Mutmap analysis suggested that Brslm was located on chromosome A07, and four candidate genes were predicted. KASP analysis demonstrated that BraA07g039390.3C was the target gene of the candidates. BraA07g039390.3C is a homologous to Arabidopsis CLV1 encoding receptor kinase with an extracellular leucine-rich domain. Sequencing analysis revealed that a single SNP from G to A occurred in 904th nucleotide of Brclv1, which resulted in the change of the 302nd amino acid from Asp to Asn. The SNP was co-segregated with the mutant phenotype in F2 individuals and located on the conserved domains. These results indicated that BrCLV1 was the mutant gene for slm which led to shortening leaves and increasing leaf numbers, disrupting the leafy heading formation in FT. These findings contribute to revealing the BrCLV1 function in leafy head formation in Chinese cabbage.
[前沿资讯 ] BASF’s vegetable seeds business and IUNU partner to advance digital phenotyping for the elevation and breeding of hydroponic lettuce varieties 进入全文
SeedQuest
近日，巴斯夫蔬菜种子事业部宣布与美国计算机视觉技术公司IUNU合作，共同推进数字表型分析，助力水培生菜育种。IUNU作为计算机视觉和人工智能软件提供商，将人工智能纳入商业温室和室内农场的检测系统，通过机器学习和计算机视觉技术，实现对个体植株健康状况大规模的跟踪和预测。该服务基于一款名为“LUNA”的人工智能系统（LUNA AI），它是由计算机视觉和人工智能软件组成的先进数字表型平台，可生成有关水培生菜生长模式和性状的数据，这将使巴斯夫能够选择合适的生菜品种进行改良和育种，从而帮助种植者提高和扩大生产规模。 巴斯夫认为，为提供高质量的种子并满足不断增长的需求，育种必须由数据驱动。此次合作是巴斯夫数字化战略的一部分，旨在通过与合作伙伴的合作来改善数据驱动的决策。通过开发和应用具有额外表型的算法，将帮助育种者提高育种预测能力，判断哪些品系在特定情况下表现最佳。
[学术文献 ] Identification of key genes controlling soluble sugar and glucosinolate biosynthesis in Chinese cabbage by integrating metabolome and genome-wide transcriptome analysis 进入全文
Front Plant Sci
Introduction: Soluble sugar and glucosinolate are essential components that determine the flavor of Chinese cabbage and consumer preferences. However, the underlying regulatory networks that modulate the biosynthesis of soluble sugar and glucosinolate in Chinese cabbage remain largely unknown. Methods: The glucosinolate and carotene content in yellow inner-leaf Chinese cabbage were observed, followed by the combination of metabolome and transcriptome analysis to explore the metabolic basis of glucosinolate and soluble sugar. Results: This study observed high glucosinolate and carotene content in yellow inner-leaf Chinese cabbage, which showed a lower soluble sugar content. The differences between the yellow and the white inner-leaf Chinese cabbage were compared using the untargeted metabonomic and transcriptomic analyses in six cultivars of Chinese cabbage to explore the metabolic basis of glucosinolate and soluble sugar. Aliphatic glucosinolate and two soluble sugars (fructose and glucose) were the key metabolites that caused the difference in Chinese cabbage’s glucosinolate and soluble sugar. By integrating soluble sugar and glucosinolate-associated metabolism and transcriptome data, we indicated BraA05gAOP1 and BraA04gAOP4, BraA03gHT7 and BraA01gHT4 were the glucosinolates and soluble sugar biosynthesis structural genes. Moreover, BraA01gCHR11 and BraA07gSCL1 were two vital transcription factors that regulate soluble sugar and glucosinolate biosynthesis. Discussion: These findings provide novel insights into glucosinolate and soluble sugar biosynthesis and a possible explanation for the significant difference in nutrients between yellow and white inner-leaf Chinese cabbage. Moreover, it will facilitate genetic modification to improve the Chinese cabbage’s nutritional and health values.
[学术文献 ] Genome-wide detection of genotype environment interactions for flowering time in Brassica napus 进入全文
Front Plant Sci
Flowering time is strongly related to the environment, while the genotype-by-environment interaction study for flowering time is lacking in Brassica napus. Here, a total of 11,700,689 single nucleotide polymorphisms in 490 B. napus accessions were used to associate with the flowering time and related climatic index in eight environments using a compressed variance-component mixed model, 3VmrMLM. As a result, 19 stable main-effect quantitative trait nucleotides (QTNs) and 32 QTN-by-environment interactions (QEIs) for flowering time were detected. Four windows of daily average temperature and precipitation were found to be climatic factors highly correlated with flowering time. Ten main-effect QTNs were found to be associated with these flowering-time-related climatic indexes. Using differentially expressed gene (DEG) analysis in semi-winter and spring oilseed rapes, 5,850 and 5,511 DEGs were found to be significantly expressed before and after vernalization. Twelve and 14 DEGs, including 7 and 9 known homologs in Arabidopsis, were found to be candidate genes for stable QTNs and QEIs for flowering time, respectively. Five DEGs were found to be candidate genes for main-effect QTNs for flowering-time-related climatic index. These candidate genes, such as BnaFLCs, BnaFTs, BnaA02.VIN3, and BnaC09.PRR7, were further validated by the haplotype, selective sweep, and co-expression networks analysis. The candidate genes identified in this study will be helpful to breed B. napus varieties adapted to particular environments with optimized flowering time.
[学术文献 ] Transcriptomic analysis of succulent stem development of Chinese kale (Brassica oleracea var. alboglabra Bailey) and its synthetic allotetraploid via RNA sequencing 进入全文
Front Plant Sci
Chinese kale (Brassica oleracea var. alboglabra Bailey, CC) is a succulent stem vegetable in the Brassica family. Its allotetraploid (AACC) vegetable germplasm, which was synthesized via distant hybridization with the colloquially named ‘yellow turnip’ (B. rapa L. ssp. rapifera Matzg., AA), has a swelling stem similar to CC. To address the molecular mechanism of stem development for CC and AACC, RNA sequencing (RNA-seq) was used to investigate transcriptional regulation of their stem development at three key stages including 28 days, 42 days and the bolting stage (BS) after sowing. As a result, 32,642, 32,665, 33,816, 32,147, 32,293 and 32,275 genes were identified in six corresponding cDNA libraries. Among them, 25,459 genes were co-expressed, while 7,183, 7,206, 8,357, 6,688, 6,834 and 6,814 genes were specifically expressed. Additionally, a total of 29,222 differentially expressed genes (DEGs) were found for functional enrichment as well as many genes involved in plant hormones including gibberellin (GA), abscisic acid (ABA), cytokinin (CTK) and auxin (AUX). Based on gene expression consistency between CC and AACC, the gene families including DELLA, GID, PYR/PYL, PP2C, A-ARR and AUX/IAA might be related to stem development. Among these, eight genes including Bo00834s040, Bo5g093140, Bo6g086770, Bo9g070200, Bo7g116570, Bo3g054410, Bo7g093470 and Bo5g136600 may play important roles in stem development based on their remarkable expression levels as confirmed by qRT-PCR. These findings provide a new theoretical basis for understanding the molecular mechanism of stem development in Brassica vegetable stem breeding.
