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1.
Int J Mol Sci ; 24(1)2023 Jan 01.
Artículo en Inglés | MEDLINE | ID: mdl-36614198

RESUMEN

Nitrate Transporter 1/Peptide Transporter Family (NPF) genes encode membrane transporters involved in the transport of diverse substrates. However, little is known about the diversity and functions of NPFs in Brassica rapa. In this study, 85 NPFs were identified in B. rapa (BrNPFs) which comprised eight subfamilies. Gene structure and conserved motif analysis suggested that BrNFPs were conserved throughout the genus. Stress and hormone-responsive cis-acting elements and transcription factor binding sites were identified in BrNPF promoters. Syntenic analysis suggested that tandem duplication contributed to the expansion of BrNPFs in B. rapa. Transcriptomic profiling analysis indicated that BrNPF2.6, BrNPF2.15, BrNPF7.6, and BrNPF8.9 were expressed in fertile floral buds, suggesting important roles in pollen development. Thirty-nine BrNPFs were responsive to low nitrate availability in shoots or roots. BrNPF2.10, BrNPF2.19, BrNPF2.3, BrNPF5.12, BrNPF5.16, BrNPF5.8, and BrNPF6.3 were only up-regulated in roots under low nitrate conditions, indicating that they play positive roles in nitrate absorption. Furthermore, many genes were identified in contrasting genotypes that responded to vernalization and clubroot disease. Our results increase understanding of BrNPFs as candidate genes for genetic improvement studies of B. rapa to promote low nitrate availability tolerance and for generating sterile male lines based on gene editing methods.


Asunto(s)
Brassica rapa , Brassica rapa/metabolismo , Nitratos/metabolismo , Perfilación de la Expresión Génica , Transportadores de Nitrato , Polen/metabolismo , Regulación de la Expresión Génica de las Plantas , Filogenia , Proteínas de Plantas/metabolismo
2.
Front Plant Sci ; 12: 734419, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-34777417

RESUMEN

Clubroot disease, which is caused by the soil-borne pathogen Plasmodiophora brassicae War (P. brassicae), is one of the oldest and most destructive diseases of Brassica and cruciferous crops in the world. Plant microRNAs [micro ribonucleic acids (miRNAs)] play important regulatory roles in several developmental processes. Although the role of plant miRNAs in plant-microbe interaction has been extensively studied, there are only few reports on the specific functions of miRNAs in response to P. brassicae. This study investigated the roles of miRNAs and their targets during P. brassicae infection in a pair of Brassica napus near-isogenic lines (NILs), namely clubroot-resistant line 409R and clubroot-susceptible line 409S. Small RNA sequencing (sRNA-seq) and degradome-seq were performed on root samples of 409R and 409S with or without P. brassicae inoculation. sRNA-seq identified a total of 48 conserved and 72 novel miRNAs, among which 18 had a significant differential expression in the root of 409R, while only one miRNA was differentially expressed in the root of 409S after P. brassicae inoculation. The degradome-seq analysis identified 938 miRNA target transcripts, which are transcription factors, enzymes, and proteins involved in multiple biological processes and most significantly enriched in the plant hormone signal transduction pathway. Between 409R and 409S, we found eight different degradation pathways in response to P. brassicae infection, such as those related to fatty acids. By combining published transcriptome data, we identified a total of six antagonistic miRNA-target pairs in 409R that are responsive to P. brassicae infection and involved in pathways associated with root development, hypersensitive cell death, and chloroplast metabolic synthesis. Our results reveal that P. brassicae infection leads to great changes in miRNA pool and target transcripts. More interestingly, these changes are different between 409R and 409S. Clarification of the crosstalk between miRNAs and their targets may shed new light on the possible mechanisms underlying the pathogen resistance against P. brassicae.

3.
Artículo en Zh | MEDLINE | ID: mdl-17167203

RESUMEN

A seed-specific Fad2 gene expression cassette, which is free-marker gene and with sense and antisense structure, was constructed by using the promoter and terminator of rape seed storage protein cruciferin gene. Transgenic rape plants without selection marker genes were obtained by Agrobacterium-mediated transformation. The oleic acid content of transgenic plant seeds is 83.9%, which is nearly the same as that of Brassica napus with double Fad2 mutation (85%). The result of RT-PCR analysis shows that the raising of oleic acid content may be due to the degradation of Fad2 mRNA induced by co-transformation of sense-antisense Fad2 gene. These transgenic plants with high oleic acid trait grew normally and without the disadvantageous agronomic traits such as weak cold resistance, tardy development, death of buds and low rate of seed setting caused by Fad2 inactivation in mutant Brassica napus plants. This work would serve as a good base for breeding of more lines with high oleic acid content.


Asunto(s)
Brassica napus/genética , Ácido Graso Desaturasas/genética , Ácido Oléico/metabolismo , Interferencia de ARN , Brassica napus/metabolismo , Ácido Graso Desaturasas/metabolismo , Regulación de la Expresión Génica de las Plantas , Vectores Genéticos/genética , Modelos Genéticos , Plantas Modificadas Genéticamente/genética , Plantas Modificadas Genéticamente/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa
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