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author:("arisaka, Mari")
1.  Breaking restricted taxonomic functionality by dual resistance genes 
Plant Signaling & Behavior  2013;8(6):e24244.
NB-LRR-type disease resistance (R) genes have been used in traditional breeding programs for crop protection. However, functional transfer of NB-LRR-type R genes to plants in taxonomically distinct families to establish pathogen resistance has not been successful. Here we demonstrate that a pair of Arabidopsis (Brassicaceae) NB-LRR-type R genes, RPS4 and RRS1, properly function in two other Brassicaceae, Brassica rapa and B. napus, but also in two Solanaceae, Nicotiana benthamiana and tomato (Solanum lycopersicum). The solanaceous plants transformed with RPS4/RRS1 confer bacterial effector-specific immunity responses. Furthermore, RPS4 and RRS1, which confer resistance to a fungal pathogen Colletotrichum higginsianum in Brassicaceae, also protect against Colletotrichum orbiculare in cucumber (Cucurbitaceae). Thus the successful transfer of two R genes at the family level overcomes restricted taxonomic functionality. This implies that the downstream components of R genes must be highly conserved and interfamily utilization of R genes can be a powerful strategy to combat pathogens.
doi:10.4161/psb.24244
PMCID: PMC3907395  PMID: 23518587
Colletotrichum higginsianum; Pseudomonas syringae; R gene; RPS4; RRS1; Ralstonia solanacearum; restricted taxonomic functionality
2.  Interfamily Transfer of Dual NB-LRR Genes Confers Resistance to Multiple Pathogens 
PLoS ONE  2013;8(2):e55954.
A major class of disease resistance (R) genes which encode nucleotide binding and leucine rich repeat (NB-LRR) proteins have been used in traditional breeding programs for crop protection. However, it has been difficult to functionally transfer NB-LRR-type R genes in taxonomically distinct families. Here we demonstrate that a pair of Arabidopsis (Brassicaceae) NB-LRR-type R genes, RPS4 and RRS1, properly function in two other Brassicaceae, Brassica rapa and Brassica napus, but also in two Solanaceae, Nicotiana benthamiana and tomato (Solanum lycopersicum). The solanaceous plants transformed with RPS4/RRS1 confer bacterial effector-specific immunity responses. Furthermore, RPS4 and RRS1, which confer resistance to a fungal pathogen Colletotrichum higginsianum in Brassicaceae, also protect against Colletotrichum orbiculare in cucumber (Cucurbitaceae). Importantly, RPS4/RRS1 transgenic plants show no autoimmune phenotypes, indicating that the NB-LRR proteins are tightly regulated. The successful transfer of two R genes at the family level implies that the downstream components of R genes are highly conserved. The functional interfamily transfer of R genes can be a powerful strategy for providing resistance to a broad range of pathogens.
doi:10.1371/journal.pone.0055954
PMCID: PMC3577827  PMID: 23437080
3.  Development of Full-Length cDNAs from Chinese Cabbage (Brassica rapa Subsp. pekinensis) and Identification of Marker Genes for Defence Response 
Arabidopsis belongs to the Brassicaceae family and plays an important role as a model plant for which researchers have developed fine-tuned genome resources. Genome sequencing projects have been initiated for other members of the Brassicaceae family. Among these projects, research on Chinese cabbage (Brassica rapa subsp. pekinensis) started early because of strong interest in this species. Here, we report the development of a library of Chinese cabbage full-length cDNA clones, the RIKEN BRC B. rapa full-length cDNA (BBRAF) resource, to accelerate research on Brassica species. We sequenced 10 000 BBRAF clones and confirmed 5476 independent clones. Most of these cDNAs showed high homology to Arabidopsis genes, but we also obtained more than 200 cDNA clones that lacked any sequence homology to Arabidopsis genes. We also successfully identified several possible candidate marker genes for plant defence responses from our analysis of the expression of the Brassica counterparts of Arabidopsis marker genes in response to salicylic acid and jasmonic acid. We compared gene expression of these markers in several Chinese cabbage cultivars. Our BBRAF cDNA resource will be publicly available from the RIKEN Bioresource Center and will help researchers to transfer Arabidopsis-related knowledge to Brassica crops.
doi:10.1093/dnares/dsr018
PMCID: PMC3158467  PMID: 21745830
Arabidopsis; Brassica rapa; full-length cDNA; jasmonic acid; salicylic acid
4.  A dual resistance gene system prevents infection by three distinct pathogens 
Plant Signaling & Behavior  2009;4(10):954-955.
Colletotrichum higginsianum causes typical anthracnose lesions on the leaves, petioles, and stems of cruciferous plants. Inoculation of Arabidopsis thaliana ecotype Columbia leaves with C. higginsianum results in fungal growth and disease symptoms reminiscent of those induced in other cruciferous plants. We performed map-based cloning and natural variation analysis of 19 A. thaliana ecotypes to identify a dominant resistance locus against C. higginsianum. We found that the A. thaliana RCH2 (for recognition of C. higginsianum) locus encodes two NB-LRR proteins, both of which are required for resistance to C. higginsianum in the A. thaliana ecotype Ws-0. Both proteins are well-characterized R proteins involved in resistance against bacterial pathogens; RRS1 (resistance to Ralstonia solanacearum 1) confers resistance to strain Rs1000 of R. solanacearum and RPS4 to Pseudomonas syringae pv. tomato strain DC3000 expressing avrRps4 (Pst-avrRps4). Furthermore, we found that both RRS1-Ws and RPS4-Ws genes are required for resistance to Pst-avrRps4 and to Rs1002 R. solanacearum. We therefore demonstrate that a pair of neighboring genes, RRS1-Ws and RPS4-Ws, function cooperatively as a dual R-gene system against at least three distinct pathogens.
PMCID: PMC2801359  PMID: 19826224
R gene; RPS4; RRS1; Colletotrichum higginsianum; Pseudomonas syringae; Ralstonia solanacearum
5.  Jasmonate-dependent plant defense restricts thrips performance and preference 
BMC Plant Biology  2009;9:97.
Background
The western flower thrips (Frankliniella occidentalis [Pergande]) is one of the most important insect herbivores of cultivated plants. However, no pesticide provides complete control of this species, and insecticide resistance has emerged around the world. We previously reported the important role of jasmonate (JA) in the plant's immediate response to thrips feeding by using an Arabidopsis leaf disc system. In this study, as the first step toward practical use of JA in thrips control, we analyzed the effect of JA-regulated Arabidopsis defense at the whole plant level on thrips behavior and life cycle at the population level over an extended period. We also studied the effectiveness of JA-regulated plant defense on thrips damage in Chinese cabbage (Brassica rapa subsp. pekinensis).
Results
Thrips oviposited more on Arabidopsis JA-insensitive coi1-1 mutants than on WT plants, and the population density of the following thrips generation increased on coi1-1 mutants. Moreover, thrips preferred coi1-1 mutants more than WT plants. Application of JA to WT plants before thrips attack decreased the thrips population. To analyze these important functions of JA in a brassica crop plant, we analyzed the expression of marker genes for JA response in B. rapa. Thrips feeding induced expression of these marker genes and significantly increased the JA content in B. rapa. Application of JA to B. rapa enhanced plant resistance to thrips, restricted oviposition, and reduced the population density of the following generation.
Conclusion
Our results indicate that the JA-regulated plant defense restricts thrips performance and preference, and plays an important role in the resistance of Arabidopsis and B. rapa to thrips damage.
doi:10.1186/1471-2229-9-97
PMCID: PMC2724403  PMID: 19635132
6.  Arabidopsis-thrips system for analysis of plant response to insect feeding 
Plant Signaling & Behavior  2008;3(7):446-447.
Insect feeding retards plant growth and decreases crop productivity. Plants respond to insect feeding at the molecular, cellular and physiological levels. The roles of the plant hormones jasmonic acid (JA), ethylene (ET) and salicylic acid (SA) in plant responses to insect feeding have been studied. However, these studies are focused on the plant responses to feeding by well-studied caterpillar type insects or aphid pests. In contrast, we have focused on a minute insect pest, the western flower thrips (Frankliniella occidentalis). Analyses of the responses of hormone-related mutants of Arabidopsis (i.e., JA-insensitive mutant coi1-1, ET-insensitive mutants ein2-1 and ein3-1, and SA-deficient mutant eds16-1) and transcriptome-based comparative analyses indicate the central role of JA in plant responses to thrips feeding. Our work clearly shows that JA signaling, but not JA/ET signaling, is involved in plant tolerance to thrips feeding. We intend to examine the utility and suitability of the Arabidopsis-thrips system in studies of plant responses to insect feeding.
PMCID: PMC2634423  PMID: 19704479
Arabidopsis thaliana; ethylene; Frankliniella occidentalis; insect feeding; jasmonate; western flower thrips

Results 1-6 (6)