Search

According to structural and functional similarities, small GTP-binding proteins can be divided into five distinct families, in which, Ras-related nuclear protein (Ran) GTPases regulates proteins an...

The fertilization of flowering plants is a critical event in their reproduction. ZnF1, a C2H2-like zinc finger protein, is highly expressed in the reproductive organs during Brassica oleracea self-pollination. However, BoZnF1’s functions during these biological processes have not been well characterized. Genetically modified mutants were employed to investigate the potential roles of ZnF1 in Arabidopsis fertilization. A Promoter expression analysis indicated that BoZnF1 is primarily expressed in root and reproductive organs, especially in filaments and pistils. When transformed into Arabidopsis protoplasts, the green fluorescent protein–BoZnF1 fusion fluoresced in the nuclei. A T-DNA insert (znf1) in Arabidopsis disrupted inflorescence development, including elongation of the gynophore and papilla cells on the stigma and the failure of anther sac cracking, resulting in defective pollen morphology. By hybrid pollination between wild-type and znf1, we found that wild-type pollen grains grew slowly on the znf1 stigma, leading to a reduced seed-setting rate. In addition, in vivo yeast two-hybrid screening and co-immunoprecipitation were performed to identify the binding protein of ZnF1. A TEOSINTE BRANCHED 1-CYCLOIDEA-PCF family transcription factor, BoTCP4, was identified as a ZnF1-interacting partner. We found that znf1 plays a role in the auxin pathway. When znf1 plants were treated with N-1-naphthylphthalamic acid, an auxin-transport inhibitor, the defects were exacerbated, leading to the ectopic formation of the gynophore in the ovarian tissue. Thus, ZnF1 may play a negative regulatory role and influence reproduction through the auxin pathway in Brassica.

Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) is not only involved in carbohydrate metabolism, but also plays an important role in stress resistance. However, it has not been reported in Brassica oleracea. In this study, we performed a genome-wide identification of BoGAPDH in B. oleracea and performed cloning and expression analysis of one of the differentially expressed genes, BoGAPC. A total of 16 members of the BoGAPDH family were identified in B. oleracea, which were conserved, distributed unevenly on chromosomes and had tandem repeat genes. Most of the genes were down-regulated during self-pollination, and the highest expression was found in stigmas and sepals. Different transcriptome data showed that BoGAPDH genes were differentially expressed under stress, which was consistent with the results of qRT-PCR. We cloned and analyzed the differentially expressed gene BoGAPC and found that it was in the down-regulated mode 1 h after self-pollination, and the expression was the highest in the stigma, which was consistent with the result of GUS staining. The promoter region of the gene not only has stress response elements and plant hormone response elements, but also has a variety of specific elements for regulating floral organ development. Subcellular localization indicates that the BoGAPC protein is located in the cytoplasm and belongs to the active protein in the cytoplasm. The results of prokaryotic expression showed that the size of the BoGAPC protein was about 37 kDa, which was consistent with the expected results, indicating that the protein was induced in prokaryotic cells. The results of yeast two-hybrid and GST pull-down showed that the SRK kinase domain interacted with the BoGAPC protein. The above results suggest that the BoGAPDH family of B. oleracea plays an important role in the process of plant stress resistance, and the BoGAPC gene may be involved in the process of self-incompatibility in B. oleracea, which may respond to SI by encoding proteins directly interacting with SRK.

Armadillo repeat containing 1 (ARC1) is phosphorylated by S-locus receptor kinase (SRK) and functions as a positive regulator in self-incompatibility response of Brassica. However, ARC1 only causes partial breakdown of the self-incompatibility response, and other SRK downstream factors may also participate in the self-incompatibility signaling pathway. In the present study, to search for SRK downstream targets, a plant U-box protein 3 (BoPUB3) was identified from the stigma of Brassica oleracea L. BoPUB3 was highly expressed in the stigma, and its expression was increased with the stigma development and reached to the highest level in the mature-stage stigma. BoPUB3, a 76.8-kDa protein with 697 amino acids, is a member of the PUB-ARM family and contains three domain characteristics of BoARC1, including a U-box N-terminal domain, a U-box motif, and a C-terminal arm repeat domain. The phylogenic tree showed that BoPUB3 was close to BoARC1. The synteny analysis revealed that B. oleracea chromosomal region containing BoPUB3 had high synteny with the Arabidopsis thaliana chromosomal region containing AtPUB3 (At3G54790). In addition, the subcellular localization analysis showed that BoPUB3 primarily localized in the plasma membrane and also in the cytoplasm. The combination of the yeast two-hybrid and in vitro binding assay showed that both BoPUB3 and BoARC1 could interact with SRK kinase domain, and SRK showed much higher level of β-galactosidase activity in its interaction with BoPUB3 than with BoARC1. These results implied that BoPUB3 is a novel interactor with SRK, which lays a basis for further research on whether PUB3 participates in the self-incompatibility signaling pathway.

Angiosperms have developed self-incompatibility (SI) systems to reject self-pollen, thereby promoting outcrossing. The Brassicaceae belongs to typical sporophytic system, having a single S-locus controlled SI response, and was chosen as a model system to study SI-related intercellular signal transduction. In this regard, the downstream factor of EXO70A1 was unknown. Here, protein two-dimensional electrophoresis (2-DE) method and coupled with matrix-assisted laser desorption ionization/time of flight of flight mass spectrometry (MALDI-TOF -MS) and peptide mass fingerprinting (PMF) was used to further explore the mechanism of SI responses in Brassica oleracea L. var. capitata L. at protein level. To further confirm the time point of protein profile change, total proteins were collected from B. oleracea pistils at 0 min, 1 h, and 2 h after self-pollination. In total 902, 1088 and 1023 protein spots were separated in 0 min, 1 h and 2 h 2-DE maps, respectively. Our analyses of self-pollination profiles indicated that proteins mainly changed at 1 h post-pollination in B. oleracea. Moreover, 1077 protein spots were separated in cross-pollinated 1 h (CP) pistil 2-DE map. MALDI-TOF-MS and PMF successfully identified 34 differentially-expressed proteins (DEPs) in SP and CP 1 h 2-DE maps. Gene ontology and KEGG analysis revealed an array of proteins grouped in the following categories: stress and defense response (35%), protein metabolism (18%), carbohydrate and energy metabolism (12%), regulation of translation (9%), pollen tube development (12%), transport (9%) and cytoskeletal (6%). Sets of DEPs identified specifically in SP or only up-regulated expressed in CP pistils were chosen for funther investigating in floral organs and during the process of self- and cross-pollination. The function of these DEPs in terms of their potential involvement in SI in B. oleracea is discussed.

The plant U-box (PUB) protein family plays an important role in plant growth and development. The U-box gene family has been well studied in Arabidopsis thaliana, Brassica rapa, rice, etc., but there have been no systematic studies in Brassica oleracea. In this study, we performed genome-wide identification and evolutionary analysis of the U-box protein family of B. oleracea. Firstly, based on the Brassica database (BRAD) and the Bolbase database, 99 Brassica oleracea PUB genes were identified and divided into seven groups (I&ndash, VII). The BoPUB genes are unevenly distributed on the nine chromosomes of B. oleracea, and there are tandem repeat genes, leading to family expansion from the A. thaliana genome to the B. oleracea genome. The protein interaction network, GO annotation, and KEGG pathway enrichment analysis indicated that the biological processes and specific functions of the BoPUB genes may mainly involve abiotic stress. RNA-seq transcriptome data of different pollination times revealed spatiotemporal expression specificity of the BoPUB genes. The differential expression profile was consistent with the results of RT-qPCR analysis. Additionally, a large number of pollen-specific cis-acting elements were found in promoters of differentially expressed genes (DEG), which verified that these significantly differentially expressed genes after self-pollination (SP) were likely to participate in the self-incompatibility (SI) process, including gene encoding ARC1, a well-known downstream protein of SI in B. oleracea. Our study provides valuable information indicating that the BoPUB genes participates not only in the abiotic stress response, but are also involved in pollination.

Self-incompatibility (SI) is an important mating system to prevent inbreeding and promote outcrossing. ARC1 and Exo70A1 function as the downstream targets of the S-locus receptor kinase and play conservative roles in Brassica SI signaling. Based on the sequence homology, Exo70A1 is divided into four subdomains: leucine zipper (Leu(128)-Leu(149)), hypervariable region (Ser(172)-Leu(197)), SUMO modification motif (Glu(260)-Ile(275)), and pfamExo70 domain (His(271)-Phe(627)). ARC1 contains four domains as follows: leucine zipper (Leu(116)-Leu(137)), coiled-coil domain (Thr(210)-Val(236)), U-box (Asp(282)-Trp(347)) motif, and ARM (Ala(415)-Thr(611)) domain. Bioinformatics analysis, yeast two-hybrid screening and pull-down assays show that leucine zipper and coiled-coil motifs of ARC1116-236 are required for the interaction with Exo70A1, while the addition of ARM motif results in loss of the interaction with Exo70A1. Meanwhile, the N-terminal of Exo70A1 without any domains shows a weak interaction with ARC1, and the level of LacZ expression increases with addition of leucine zipper and reaches the maximum value with hypervariable region and SUMO modification motif, indicating that hypervariable region and SUMO modification motif of Exo70A1172-275 is mainly responsible for the binding with ARC1, whereas pfamExo70 domain has little affinity for ARC1. Lys(181) located in the Exo70A1 hypervariable region may be the ubiquitination site mediating the interaction between ARC1 and Exo70A1. Therefore, both the leucine zipper with coiled-coil structure of ARC1116-236, and the hypervariable region and SUMO modification motif of Exo70A1172-275 are the core interaction domains between ARC1 and Exo70A1. Any factors affecting these core domains would be the regulators of ARC1 mediating ubiquitin degradation in self-incompatible system.

In order to identify the functional domains which regulate the interaction between the self-incompatibility proteins armadillo repeat containing 1 (ARC1) and exocyst 70 A1 (Exo70A1) in Brassica oleracea, fragments containing selected motifs of ARC1 (ARC1210, ARC1246, ARC1279, ARC1354) and site-specific mutants with substitutions at possible interaction sites (ARC1354m, ARC1664m) were PCR amplified and inserted into pGADT7, while coding sequences from Exo70A1 (Exo70A185, Exo70A1) were subcloned into pGBKT7. The interactions between the protein products produced by these constructs were then analyzed utilizing a yeast two-hybrid system. Our data indicate that both ARC1210 and ARC1246 interact strongly with Exo70A185 and Exo70A1, while ARC1279, ARC1354, ARC1354m and ARC1664m exhibited a weak interaction, indicating that the recognition sites are located within the 210 N-terminal amino acids of ARC1 and the 85 N-terminal amino acids of Exo70A1. This was further verified by GST pull-down analysis. This supports a model in which the N-terminal leucine zipper of ARC1 and the first 85 N-terminal amino acids of Exo70A1 mediate the interaction between these two proteins. Bioinformatic and phylogenetic analysis demonstrated that these motifs were highly conserved across different species, indicating that the interaction characterized in B. oleracea may operate in a wide array of cultivars.

The low resolution of chromosome-based Fluorescence in situ hybridisation (FISH) mapping is primarily due to the structure of the plant cell wall and cytoplasm and the compactness of regular chromosomes, which represent a significant obstacle to FISH. In order to improve spatial resolution and signal detection sensitivity, we provide a reproducible method to generate high-quality extended chromosomes that are ~13 times as long as their pachytene counterparts. We demonstrate that proteinase K used in this procedure is crucial for stretching pachytene chromosomes of Brassica oleracea in the context of a modified Carnoy's II fixative (6:1:3, ethanol:chloroform:acetic acid). The quality of super-stretched chromosomes was assessed in several FISH experiments. FISH signals from both repetitive 5S rDNA and single-copy ARC1 on super-stretched chromosomes are brighter than those on other different types of chromosome due to enhanced accessibility to targets on stretched pachytene chromosomes. In conclusion, the resulting extended chromosomes are suitable for FISH mapping for repetitive DNA sequences and the localisation of a single-copy locus, and FISH performed on super-stretched chromosomes can achieve significantly higher sensitivity and spatial resolution than other chromosome-based FISH mapping techniques.

从甘蓝、大白菜与甘蓝型油菜中分离出 EXO70A1 基因，对该基因的序列进行生物信息学分析, 然后转化酵母Y187, 应用半定量RT-PCR检测 BoEXO70A1 基因的表达特性。结果表明, 3种芸薹属植物 EXO70A1 序列长度均为1 917 bp，相似性97.1%, 它们的gDNA序列均为单一序列，长度分别为3 797、3 752和3 770 bp，一致度达91.0%，均由12个外显子及11个内含子组成，除了第4、第5、第6、第8个内含子外，其余内含子的保守性低于外显子; 推导的3种蛋白质序列(BnEXO70A1、BrEXO70A1和BoEXO70A1)的相似度与一致性分别达99.8%与98.1%，其二级结构、三维结构及理化特性高度相似。 EXO70A1 基因的11个内含子的剪切位点均符合“GU-AG”法则，剪切受体(AG)的前20~50个碱基存在一段保守的序列“CU(A/G)A(C/U)”; 3种芸薹属植物与拟南芥 EXO70A1 基因的12个外显子的对应序列长度完全相同，所构成的编码区的序列一致性达90.1%，相应的蛋白质序列的相似度与一致性分别达99.8%与93.7%; 分子进化分析表明, EXO70A1在整个EXO70蛋白家族中及不同的植物间表现出较高的保守性; BoEXO70A1 在酵母细胞Y187呈现弱表达; EXO70A1 在甘蓝的雄蕊、幼茎、幼嫩花瓣、雌蕊、幼根及叶片中均能表达，可能属于组成型表达基因，但是其表达量在不同发育时期的不同器官中存在差异，授粉前雌蕊中最高，雄蕊中最低。由此可知， EXO70A1 在芸薹属植物中整体高度保守, 但在酵母转化株和甘蓝各器官中的组成型表达有所差异，推测 EXO70A1 在植物细胞中具有多种重要的功能。

Understanding the synergic interactions between arbuscular mycorrhizal fungi (AMF) and its host mulberry (Morus alba L.), an important perennial multipurpose plant, has theoretical and practical significance in mulberry plantation, silkworm cultivation and relevant textile industry. In a greenhouse study, we compared functional distinctions of three genetically different AMF species (Acaulospora scrobiculata, Funneliformis mosseae and Rhizophagus intraradices) on physiological and growth characteristics as well as leaf quality of 6-month-old mulberry seedlings. Results showed that mulberry was AMF-species dependent, and AMF colonization significantly increased shoot height and taproot length, stem base and taproot diameter, leaf and fibrous root numbers, and shoot and root biomass production. Meanwhile, leaf chlorophyll a or b and carotenoid concentrations, net photosynthetic rate, transpiration rate and stomatal conductance were generally significantly greater, while intercellular CO2 concentration was significantly lower in AMF-inoculated seedlings than in non-AMF-inoculated counterparts. These trends were also generally true for leaf moisture, total nitrogen, all essential amino acids, histidine, proline, soluble protein, sugar and fatty acid as they were significantly increased under mycorrhization. Among these three tested AMFs, significantly greater effects of AMF on above-mentioned mulberry physiological and growth characteristics ranked as F. mosseae > A. scrobiculata > R. intraradices, whilst on mulberry leaf quality (e.g. nutraceutical values) for better silkworm growth as F. mosseae ≈ A. scrobiculata > R. intraradices. In conclusion, our results showed that greater mulberry biomass production, and nutritional quality varied with AMF species or was AMF-species dependent. Such improvements were mainly attributed to AMF-induced positive alterations of mulberry leaf photosynthetic pigments, net photosynthetic rate, transpiration rate and N-containing compounds (methionine, threonine, histidine and proline). As a result, application of Funneliformis mosseae or Acaulospora scrobiculata in mulberry plantation could be a promising management strategy to promote silkworm cultivation and relevant textile industry.

M locus protein kinase, one of the SRK-interacting proteins, is a necessary positive regulator for the self-incompatibility response in Brassica. In B. rapa, MLPK is expressed as two different transcripts, MLPKf1 and MLPKf2, and either isoform can complement the mlpk/mlpk mutation. The AtAPK1B gene has been considered to be the ortholog of BrMLPK, and AtAPK1B has no role in self-incompatibility (SI) response in A. thaliana SRK-SCR plants. Until now, what causes the MLPK and APK1B function difference during SI response in Brassica and A. thaliana SRKb-SCRb plants has remained unknown. Here, in addition to the reported MLPKf1/2, we identified the new MLPKf1 homologous gene MLPKn1 from B. oleracea. BoMLPKn1 and BoMLPKf1 shared nucleotide sequence identity as high as 84.3 %, and the most striking difference consisted in two fragment insertions in BoMLPKn1. BoMLPKn1 and BoMLPKf1 had a similar gene structure; both their deduced amino acid sequences contained a typical plant myristoylation consensus sequence and a Ser/Thr protein kinase domain. BoMLPKn1 was widely expressed in petal, sepal, anther, stigma and leaf. Genome-wide survey revealed that the B. oleracea genome contained three MLPK homologous genes: BoMLPKf1/2, BoMLPKn1 and Bol008343n. The B. rapa genome also contained three MLPK homologous genes, BrMLPKf1/2, BraMLPKn1 and Bra040929. Phylogenetic analysis revealed that BoMLPKf1/2 and BrMLPKf1/2 were phylogenetically more distant from AtAPK1A than Bol008343n, Bra040929, BraMLPKn1 and BoMLPKn1, Synteny analysis revealed that the B. oleracea chromosomal region containing BoMLPKn1 displayed high synteny with the A. thaliana chromosomal region containing APK1B, whereas the B. rapa chromosomal region containing BraMLPKn1 showed high synteny with the A. thaliana chromosomal region containing APK1B. Together, these results revealed that BoMLPKn1/BraMLPKn1, and not the formerly reported BoMLPKf1/2 (BrMLPKf1/2), was the orthologous genes of AtAPK1B, and no ortholog of BoMLPKf1/2 (BrMLPKf1/2) was found in the A. thaliana genome. We speculated that Brassica MLPKf1/2 might have emerged after speciation of Brassica and A. thailiana, and that it was recruited to the SRK-triggered SI signaling cascade in Brassica.

ARC1 and EXO70A1 are important signal elements of self-incompatibility in Brassica . To characterize the interaction of ARC1-EXO70A1 during the course of self-incompatibility, the coding sequences of ARC1 and EXO70A1 were cloned from Brassica napus L. and B. oleracea L. var. acephala . Sequence analysis showed that ARC1 consisted of 663 amino acids in B. oleracea and 661 amino acids in B. napus , with a 45-amino-acid difference between them. Sequence alignment showed 95.9% similarity, with 93.9% exact match between BoARC1 and BnARC1. Only a 6-amino-acid difference was detected between BoEXO70A1 and BnEXO70A1, with 99.4% similarity and 98.9% exact match according to further sequence alignment. The homology between EXO70A1 alleles was higher than that between ARC1 alleles. Yeast 2-hybrid results indicated that a strong interaction existed between ARC1 and EXO70A1, which could activate the expressions of 4 reporter genes ( ADE2 , HIS3 , AUR1-C , and MEL1 ) in diploid yeast. However, there was very weak interaction between EXO70A1 and a 316-C-terminal-deletion mutant of ARC1 (ARC1 N ), which only activated the expressions from 3 reporter genes ( ADE2 , AUR1-C , and MEL1 ). This indicated that the interaction interface between ARC1 and EXO70A1 might not reside within the Armadillo (ARM) repeat domains of ARC1. The N-terminal domains of ARC1 play an essential role in the interaction of ARC1 with EXO70A1. The influence of the differences in amino-acid composition between BoARC1 and BnARC1 on the interaction between ARC1-EXO70A1 was not detected with a yeast 2-hybrid system, which may indicate that the binding interface between ARC1 and EXO70A1 was not altered by sequence differences between the 2 proteins in these Brassica species.

ARC1 is an arm repeat containing a downstream gene of S-locus receptor kinase (SRK). It interacts with SRK probably in signal transduction pathway of self-incompatibility. In this study, the interaction between SRK and ARC1 proteins was tested using the expression vectors pET43.1a-ARC1 and pET43.1a-SRK E1 . The coding sequences of ARC1 gene were amplified from stigma mRNA of Brassica oleracea L. and inserted into the expression vector pET43.1a to construct the recombinant plasma pET43.1a-ARC1. The recombinant plasmid was transformed to Escherichia coli (BL21) and the expression of the recombinant protein was detected using sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Using the co-immunoprecipitation theory and characteristic of 6× His Tag combined with Ni+ in pET43.1a-ARC1, a new method was proposed for testing the interaction between proteins. With this method the interaction between ARC1 and SRK was analyzed, showing that ARC1 and SRK could interact with each other to combine and form a complex. This study provides the theoretical and technical bases for further analyzing the mechanism of interaction between ARC1 and SRK proteins and for probing into interaction between ARC1 and downstream targets.

To understand the physical locations of self-incompatibility (SI) genes and their linkage relationship, this study conducted fluorescence in situ hybridization (FISH) in Brassica oleracea genome using the probes of MLPK and SSP genes on prometaphase chromosomes, early pachytene chromosomes, and extended DNA fibers. The results indicated that the MLPK probe was hybridized onto the short arm of a pair of homologous prometaphase chromosomes, and the average percent distance from the centromere to the signal point was 53.41±3.16. The SSP probe was hybridized onto the long arm of a pair of homologous prometaphase chromosomes with the satellite, and the average percent distance from the centromere to the signal point was 78.36±4.26. Hybridization signals from the 3 types of cytological targets with different FISH resolutions showed that both MLPK and SSP might be located at a single-copy locus in B. oleracea genome. Repeated FISH analysis indicated that both MLPK and 5S rDNA probes were hybridized onto the same chromosomes. According to the karyotype standard, MLPK is primarily inferred on chromosome 2, and SSP on chromosome 7. The results presumably revealed that neither MLPK nor SSP should be linked to S-locus, and they were located on different chromosomes of B. oleracea.

The S -locus receptor kinase (SRK) and thioredoxin-like protein 1 (THL1) from Brassica oleracea L. probably interact with each other in self-incompatibility signal process. To testify the interaction, the coding sequences of SRK kinase domain (termed SRK E1 ) and THL1 were amplified from stigma mRNA of B. oleracea . They were, then, inserted into the expression vector pET43.1a and yeast expression vector pPIC9K to construct the recombinant plasma pET43.1a-SRK E1 , pPIC9K-SRK E1 , and pET43.1a-THL1. The recombinant proteins were expressed in Escherichia coli (BL21) and Pichia pastoris (GS115), respectively. The recombinant THL1 was approved with oxidation/reduction activity by testing the ability of THL1 reducing insulin. Using a new method for testing the interaction between proteins, SRK E1 and THL1 were identified to combine with each other and form a stable complex.

The compactness of plant chromosome and the structures of plant cell wall and cytoplasm pose a great resistance to fluorescence in situ hybridization (FISH), and consequently many new methods for improving spatial resolution are being exploited to overcome these problems. However, for plants with small chromosomes like rice and Brassica, there are still many difficulties. In this article a new and effective technique for preparation of extended DNA fibers (EDFs), using a series of treatments to prophase I chromosomes of Brassica oleracea PMCs, is presented. This technique allows longitudinal extension of the chromosomes 30-107 times longer than those of their metaphase counterparts. The length of the extended DNA fibers is between 89 microm and 273 microm, and the space resolution is 42.8-53.0 kb. Stretching ratios were assessed in a number of FISH experiments with super-stretched chromosomes from meiotic prophase I nuclei of B. olerecea. Through FISH to EDFs of pachytene chromosomes hybridized in situ with SRK (S-locus receptor kinase) and SPII (S-locus protein II) probes, for the first time we localized the accurate positions of S-locus and quantitatively analyzed the features of S genes in B. oleracea genome to show all S genes were single-copied. In addition, the length between two linked genes was measured to be about one micron. As a result, the highest space resolution which was about 4 kb was obtained.

The compactness of plant chromosomes and the structure of the plant cell wall and cytoplasm provide a great obstacle to fluorescence in situ hybridization (FISH) for single-copy or low-copy DNA sequences. Consequently, many new methods for improving spatial resolution via chromosomal stretching have been employed to overcome this technical challenge. In this article, a technique for extracting cell-wall free nuclei at mitotic interphase, then using these nuclei to prepare extended DNA fibers (EDFs) by the method of a receding interface, whereby slide-mounted chromatin produces EDFs in concert with gravity-assisted buffer flow, was adopted as a result of the low frequency of EDF damage produced by this procedure. To examine the quality of these EDFs, we used single-copy gene encoding S-locus receptor kinase and multi-copy 5S rDNA (ribosomal DNA) as probes. The resulting EDFs proved suitable for high-resolution FISH mapping for repetitive DNA sequences, and the localization of a single-copy locus.

In recent years certain progress in Brassica signaling was reviewed about some self-compatibility-related genes such as SRK, SLG, SCR, ARC1, THL1 and THL2. Meanwhile, molecular mechanism in Brassica self-compatibility signaling was reviewed, including its action models.