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Honghwa Chen - One of the best experts on this subject based on the ideXlab platform.
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Terpene Synthase-b and Terpene Synthase-e/f Genes Produce Monoterpenes for Phalaenopsis bellina Floral Scent
'Frontiers Media SA', 2021Co-Authors: Hsin Huang, Yu Chen Chuang, Honghwa Chen, Wen Huei Chen, Yi-wei Kuo, Ya-ping Yang, Li-min Huang, Mei-fen JengAbstract:Orchids are the most species-rich plants and highly interactive with pollinators via visual or olfactory cues. Biosynthesis and emission of volatile organic compounds (VOCs) to the atmosphere facilitate the olfactory cues and ensure successful pollination. Phalaenopsis bellina is a scented orchid with monoterpenes as major VOCs, comprising linalool, geraniol, and their derivatives. Comparative transcriptomics analysis identified four terpene synthase-b (TPS-b) genes and two TPS-e/f genes with differential gene expression between scented and scentless Phalaenopsis species. Here, we confirmed their differential expression between scented and scentless Phalaenopsis orchids and excluded one TPS-b candidate. We analyzed the temporal and spatial expression and functionally characterized these TPSs. Both TPS-b and TPS-e/f genes showed an increased expression on blooming day or 3 days post-anthesis (D + 3) before the optimal emission of floral scent on D + 5, with especially high expression of PbTPS5 and PbTPS10. The TPS-b genes are expressed exclusively in reproductive organs, whereas the TPS-e/f genes are expressed in both reproductive and vegetative organs. In planta functional characterization of both PbTPS5 and PbTPS10 in tobacco and scentless Phalaenopsis plants did not produce terpenoids. Further ectopic expression in scented Phalaenopsis cultivar P. I-Hsin Venus showed that linalool was the main product, with PbTPS10 displaying 3-fold higher activity than PbTPS5. On in vitro enzyme assay with purified recombinant TPS-b proteins ectopically expressed in Escherichia coli, geraniol was the product catalyzed by PbTPS5 and PbTPS9. PbTPS3 was a linalool/(β)-cis-ocimene synthase and PbTPS4 a linalool synthase. In conclusion, both TPS-b and TPS-e/f enzymes orchestrated floral monoterpene biosynthesis in P. bellina
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PePIF1, a P-lineage of PIF-like transposable element identified in protocorm-like bodies of Phalaenopsis orchids
BMC, 2019Co-Authors: Chia-chi Hsu, Wen Chieh Tsai, Wen Huei Chen, Yu Yun Hsiao, Tien Chih Chen, Pei-han Lai, Jui-lin Hsu, Ching-hsiu Tsai, Honghwa ChenAbstract:Abstract Background Orchids produce a colorless protocorm by symbiosis with fungi upon seed germination. For mass production of orchids, the prevailing approaches are both generation of protocorm-like bodies (PLBs) from callus and multiplication of adventitious buds on inflorescence. However, somaclonal variations occur during micropropagation. Results We isolated the two most expressed transposable elements belonging to P Instability Factor (PIF)-like transposons. Among them, a potential autonomous element was identified by similarity analysis against the whole-genome sequence of Phalaenopsis equestris and named PePIF1. It contains a 19-bp terminal inverted repeat flanked by a 3-bp target site duplication and two coding regions encoding ORF1- and transposase-like proteins. Phylogenetic analysis revealed that PePIF1 belongs to a new P-lineage of PIF. Furthermore, two distinct families, PePIF1a and PePIF1b, with 29 and 37 putative autonomous elements, respectively, were isolated, along with more than 3000 non-autonomous and miniature inverted-repeat transposable element (MITE)-like elements. Among them, 828 PePIF1-related elements were inserted in 771 predicted genes. Intriguingly, PePIF1 was transposed in the somaclonal variants of Phalaenopsis cultivars, as revealed by transposon display, and the newly inserted genes were identified and sequenced. Conclusion A PIF-like element, PePIF1, was identified in the Phalaenopsis genome and actively transposed during micropropagation. With the identification of PePIF1, we have more understanding of the Phalaenopsis genome structure and somaclonal variations during micropropagation for use in orchid breeding and production
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Additional file 2: of PePIF1, a P-lineage of PIF-like transposable element identified in protocorm-like bodies of Phalaenopsis orchids
2019Co-Authors: Chia-chi Hsu, Wen Chieh Tsai, Wen Huei Chen, Yu Yun Hsiao, Tien Chih Chen, Pei-han Lai, Jui-lin Hsu, Ching-hsiu Tsai, Honghwa ChenAbstract:Table S1. Mapping of the three transposable element candidates to the whole-genome sequence of Phalaenopsis equestris. (DOCX 13 kb
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a dual repeat cis element determines expression of geranyl diphosphate synthase for monoterpene production in Phalaenopsis orchids
Frontiers in Plant Science, 2018Co-Authors: Yu Chen Chuang, Wen Chieh Tsai, Yi Chu Hung, Masaru Ohmetakagi, Wen Huei Chen, Nobutaka Mitsuda, Honghwa ChenAbstract:Phalaenopsis bellina is a scented orchid emitting large amount of monoterpenes. GERANYL DIPHOSPHATE SYNTHASE (PbGDPS) is the key enzyme for monoterpene biosynthesis, and shows concomitant expression with the emission of monoterpenes during flower development in P. bellina. Here, we identified a dual repeat cis-element in the GDPS promoter that is critical for monoterpene biosynthesis in Phalaenopsis orchids. A strong correlation between the dual repeat and the monoterpene production was revealed by examination of the GDPS promoter fragments over 12 Phalaenopsis species. Serial-deletion of the 2-kb GDPS promoter fragments demonstrated that the integrity of the dual repeat was crucial for its promoter activities. By screening the Arabidopsis transcription factors (TFs) cDNA library using yeast one-hybrid assay, AtbZIP18, a member of group I of bZIP TFs, was identified to be able to bind the dual repeat. We then identified PbbZIP4 in the transcriptome of P. bellina, showing 83% identity in the DNA binding region with that of AtbZIP18, and the expression level of PbbZIP4 was higher in the scented orchids. In addition, PbbZIP4 transactivated the GDPS promoter fragment containing the dual repeat in dual luciferase assay. Furthermore, transient ectopic expression of PbbZIP4 induced a 10-fold production of monoterpenoids in the scentless orchid. In conclusion, these results indicate that the dual repeat is a real TF-bound cis-element significant for GDPS gene expression, and thus subsequent monoterpene biosynthesis in the scented Phalaenopsis orchids.
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flower development of Phalaenopsis orchid involves functionally divergent sepallata like genes
New Phytologist, 2014Co-Authors: Zhao Jun Pan, Wen Chieh Tsai, You Yi Chen, Yun Yu Chen, Meichu Chung, Chunneng Wang, Honghwa ChenAbstract:Summary The Phalaenopsis orchid produces complex flowers that are commercially valuable, which has promoted the study of its flower development. E-class MADS-box genes, SEPALLATA (SEP), combined with B-, C- and D-class MADS-box genes, are involved in various aspects of plant development, such as floral meristem determination, organ identity, fruit maturation, seed formation and plant architecture. Four SEP-like genes were cloned from Phalaenopsis orchid, and the duplicated PeSEPs were grouped into PeSEP1/3 and PeSEP2/4. All PeSEPs were expressed in all floral organs. PeSEP2 expression was detectable in vegetative tissues. The study of protein–protein interactions suggested that PeSEPs may form higher order complexes with the B-, C-, D-class and AGAMOUS LIKE6-related MADS-box proteins to determine floral organ identity. The tepal became a leaf-like organ when PeSEP3 was silenced by virus-induced silencing, with alterations in epidermis identity and contents of anthocyanin and chlorophyll. Silencing of PeSEP2 had minor effects on the floral phenotype. Silencing of the E-class genes PeSEP2 and PeSEP3 resulted in the downregulation of B-class PeMADS2-6 genes, which indicates an association of PeSEP functions and B-class gene expression. These findings reveal the important roles of PeSEP in Phalaenopsis floral organ formation throughout the developmental process by the formation of various multiple protein complexes.
Wen Chieh Tsai - One of the best experts on this subject based on the ideXlab platform.
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the ancestral duplicated dl crc orthologs pedl1 and pedl2 function in orchid reproductive organ innovation
Journal of Experimental Botany, 2021Co-Authors: You Yi Chen, Nobutaka Mitsuda, Yu Yun Hsiao, Chuan Ming Yeh, Hongxing Yang, Chi Chou Chiu, Song Bin Chang, Zhongjian Liu, Wen Chieh TsaiAbstract:Orchid gynostemium, the fused organ of the androecium and gynoecium, and ovule development are unique developmental processes. Two DROOPING LEAF/CRABS CLAW (DL/CRC) genes, PeDL1 and PeDL2, were identified from the Phalaenopsis orchid genome and functionally characterized. Phylogenetic analysis indicated that the most recent common ancestor of orchids contained the duplicated DL/CRC-like genes. Temporal and spatial expression analysis indicated that PeDL genes are specifically expressed in the gynostemium and at the early stages of ovule development. Both PeDLs could partially complement an Arabidopsis crc-1 mutant. Virus-induced gene silencing (VIGS) of PeDL1 and PeDL2 affected the number of protuberant ovule initials differentiated from the placenta. Transient overexpression of PeDL1 in Phalaenopsis orchids caused abnormal development of ovule and stigmatic cavity of gynostemium. PeDL1, but not PeDL2, could form a heterodimer with Phalaenopsis equestris CINCINNATA 8 (PeCIN8). Paralogous retention and subsequent divergence of the gene sequences of PeDL1 and PeDL2 in P. equestris might result in the differentiation of function and protein behaviors. These results reveal that the ancestral duplicated DL/CRC-like genes play important roles in orchid reproductive organ innovation.
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PePIF1, a P-lineage of PIF-like transposable element identified in protocorm-like bodies of Phalaenopsis orchids
BMC, 2019Co-Authors: Chia-chi Hsu, Wen Chieh Tsai, Wen Huei Chen, Yu Yun Hsiao, Tien Chih Chen, Pei-han Lai, Jui-lin Hsu, Ching-hsiu Tsai, Honghwa ChenAbstract:Abstract Background Orchids produce a colorless protocorm by symbiosis with fungi upon seed germination. For mass production of orchids, the prevailing approaches are both generation of protocorm-like bodies (PLBs) from callus and multiplication of adventitious buds on inflorescence. However, somaclonal variations occur during micropropagation. Results We isolated the two most expressed transposable elements belonging to P Instability Factor (PIF)-like transposons. Among them, a potential autonomous element was identified by similarity analysis against the whole-genome sequence of Phalaenopsis equestris and named PePIF1. It contains a 19-bp terminal inverted repeat flanked by a 3-bp target site duplication and two coding regions encoding ORF1- and transposase-like proteins. Phylogenetic analysis revealed that PePIF1 belongs to a new P-lineage of PIF. Furthermore, two distinct families, PePIF1a and PePIF1b, with 29 and 37 putative autonomous elements, respectively, were isolated, along with more than 3000 non-autonomous and miniature inverted-repeat transposable element (MITE)-like elements. Among them, 828 PePIF1-related elements were inserted in 771 predicted genes. Intriguingly, PePIF1 was transposed in the somaclonal variants of Phalaenopsis cultivars, as revealed by transposon display, and the newly inserted genes were identified and sequenced. Conclusion A PIF-like element, PePIF1, was identified in the Phalaenopsis genome and actively transposed during micropropagation. With the identification of PePIF1, we have more understanding of the Phalaenopsis genome structure and somaclonal variations during micropropagation for use in orchid breeding and production
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Additional file 2: of PePIF1, a P-lineage of PIF-like transposable element identified in protocorm-like bodies of Phalaenopsis orchids
2019Co-Authors: Chia-chi Hsu, Wen Chieh Tsai, Wen Huei Chen, Yu Yun Hsiao, Tien Chih Chen, Pei-han Lai, Jui-lin Hsu, Ching-hsiu Tsai, Honghwa ChenAbstract:Table S1. Mapping of the three transposable element candidates to the whole-genome sequence of Phalaenopsis equestris. (DOCX 13 kb
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a dual repeat cis element determines expression of geranyl diphosphate synthase for monoterpene production in Phalaenopsis orchids
Frontiers in Plant Science, 2018Co-Authors: Yu Chen Chuang, Wen Chieh Tsai, Yi Chu Hung, Masaru Ohmetakagi, Wen Huei Chen, Nobutaka Mitsuda, Honghwa ChenAbstract:Phalaenopsis bellina is a scented orchid emitting large amount of monoterpenes. GERANYL DIPHOSPHATE SYNTHASE (PbGDPS) is the key enzyme for monoterpene biosynthesis, and shows concomitant expression with the emission of monoterpenes during flower development in P. bellina. Here, we identified a dual repeat cis-element in the GDPS promoter that is critical for monoterpene biosynthesis in Phalaenopsis orchids. A strong correlation between the dual repeat and the monoterpene production was revealed by examination of the GDPS promoter fragments over 12 Phalaenopsis species. Serial-deletion of the 2-kb GDPS promoter fragments demonstrated that the integrity of the dual repeat was crucial for its promoter activities. By screening the Arabidopsis transcription factors (TFs) cDNA library using yeast one-hybrid assay, AtbZIP18, a member of group I of bZIP TFs, was identified to be able to bind the dual repeat. We then identified PbbZIP4 in the transcriptome of P. bellina, showing 83% identity in the DNA binding region with that of AtbZIP18, and the expression level of PbbZIP4 was higher in the scented orchids. In addition, PbbZIP4 transactivated the GDPS promoter fragment containing the dual repeat in dual luciferase assay. Furthermore, transient ectopic expression of PbbZIP4 induced a 10-fold production of monoterpenoids in the scentless orchid. In conclusion, these results indicate that the dual repeat is a real TF-bound cis-element significant for GDPS gene expression, and thus subsequent monoterpene biosynthesis in the scented Phalaenopsis orchids.
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flower development of Phalaenopsis orchid involves functionally divergent sepallata like genes
New Phytologist, 2014Co-Authors: Zhao Jun Pan, Wen Chieh Tsai, You Yi Chen, Yun Yu Chen, Meichu Chung, Chunneng Wang, Honghwa ChenAbstract:Summary The Phalaenopsis orchid produces complex flowers that are commercially valuable, which has promoted the study of its flower development. E-class MADS-box genes, SEPALLATA (SEP), combined with B-, C- and D-class MADS-box genes, are involved in various aspects of plant development, such as floral meristem determination, organ identity, fruit maturation, seed formation and plant architecture. Four SEP-like genes were cloned from Phalaenopsis orchid, and the duplicated PeSEPs were grouped into PeSEP1/3 and PeSEP2/4. All PeSEPs were expressed in all floral organs. PeSEP2 expression was detectable in vegetative tissues. The study of protein–protein interactions suggested that PeSEPs may form higher order complexes with the B-, C-, D-class and AGAMOUS LIKE6-related MADS-box proteins to determine floral organ identity. The tepal became a leaf-like organ when PeSEP3 was silenced by virus-induced silencing, with alterations in epidermis identity and contents of anthocyanin and chlorophyll. Silencing of PeSEP2 had minor effects on the floral phenotype. Silencing of the E-class genes PeSEP2 and PeSEP3 resulted in the downregulation of B-class PeMADS2-6 genes, which indicates an association of PeSEP functions and B-class gene expression. These findings reveal the important roles of PeSEP in Phalaenopsis floral organ formation throughout the developmental process by the formation of various multiple protein complexes.
Wen Huei Chen - One of the best experts on this subject based on the ideXlab platform.
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Terpene Synthase-b and Terpene Synthase-e/f Genes Produce Monoterpenes for Phalaenopsis bellina Floral Scent
'Frontiers Media SA', 2021Co-Authors: Hsin Huang, Yu Chen Chuang, Honghwa Chen, Wen Huei Chen, Yi-wei Kuo, Ya-ping Yang, Li-min Huang, Mei-fen JengAbstract:Orchids are the most species-rich plants and highly interactive with pollinators via visual or olfactory cues. Biosynthesis and emission of volatile organic compounds (VOCs) to the atmosphere facilitate the olfactory cues and ensure successful pollination. Phalaenopsis bellina is a scented orchid with monoterpenes as major VOCs, comprising linalool, geraniol, and their derivatives. Comparative transcriptomics analysis identified four terpene synthase-b (TPS-b) genes and two TPS-e/f genes with differential gene expression between scented and scentless Phalaenopsis species. Here, we confirmed their differential expression between scented and scentless Phalaenopsis orchids and excluded one TPS-b candidate. We analyzed the temporal and spatial expression and functionally characterized these TPSs. Both TPS-b and TPS-e/f genes showed an increased expression on blooming day or 3 days post-anthesis (D + 3) before the optimal emission of floral scent on D + 5, with especially high expression of PbTPS5 and PbTPS10. The TPS-b genes are expressed exclusively in reproductive organs, whereas the TPS-e/f genes are expressed in both reproductive and vegetative organs. In planta functional characterization of both PbTPS5 and PbTPS10 in tobacco and scentless Phalaenopsis plants did not produce terpenoids. Further ectopic expression in scented Phalaenopsis cultivar P. I-Hsin Venus showed that linalool was the main product, with PbTPS10 displaying 3-fold higher activity than PbTPS5. On in vitro enzyme assay with purified recombinant TPS-b proteins ectopically expressed in Escherichia coli, geraniol was the product catalyzed by PbTPS5 and PbTPS9. PbTPS3 was a linalool/(β)-cis-ocimene synthase and PbTPS4 a linalool synthase. In conclusion, both TPS-b and TPS-e/f enzymes orchestrated floral monoterpene biosynthesis in P. bellina
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PePIF1, a P-lineage of PIF-like transposable element identified in protocorm-like bodies of Phalaenopsis orchids
BMC, 2019Co-Authors: Chia-chi Hsu, Wen Chieh Tsai, Wen Huei Chen, Yu Yun Hsiao, Tien Chih Chen, Pei-han Lai, Jui-lin Hsu, Ching-hsiu Tsai, Honghwa ChenAbstract:Abstract Background Orchids produce a colorless protocorm by symbiosis with fungi upon seed germination. For mass production of orchids, the prevailing approaches are both generation of protocorm-like bodies (PLBs) from callus and multiplication of adventitious buds on inflorescence. However, somaclonal variations occur during micropropagation. Results We isolated the two most expressed transposable elements belonging to P Instability Factor (PIF)-like transposons. Among them, a potential autonomous element was identified by similarity analysis against the whole-genome sequence of Phalaenopsis equestris and named PePIF1. It contains a 19-bp terminal inverted repeat flanked by a 3-bp target site duplication and two coding regions encoding ORF1- and transposase-like proteins. Phylogenetic analysis revealed that PePIF1 belongs to a new P-lineage of PIF. Furthermore, two distinct families, PePIF1a and PePIF1b, with 29 and 37 putative autonomous elements, respectively, were isolated, along with more than 3000 non-autonomous and miniature inverted-repeat transposable element (MITE)-like elements. Among them, 828 PePIF1-related elements were inserted in 771 predicted genes. Intriguingly, PePIF1 was transposed in the somaclonal variants of Phalaenopsis cultivars, as revealed by transposon display, and the newly inserted genes were identified and sequenced. Conclusion A PIF-like element, PePIF1, was identified in the Phalaenopsis genome and actively transposed during micropropagation. With the identification of PePIF1, we have more understanding of the Phalaenopsis genome structure and somaclonal variations during micropropagation for use in orchid breeding and production
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Additional file 2: of PePIF1, a P-lineage of PIF-like transposable element identified in protocorm-like bodies of Phalaenopsis orchids
2019Co-Authors: Chia-chi Hsu, Wen Chieh Tsai, Wen Huei Chen, Yu Yun Hsiao, Tien Chih Chen, Pei-han Lai, Jui-lin Hsu, Ching-hsiu Tsai, Honghwa ChenAbstract:Table S1. Mapping of the three transposable element candidates to the whole-genome sequence of Phalaenopsis equestris. (DOCX 13 kb
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a dual repeat cis element determines expression of geranyl diphosphate synthase for monoterpene production in Phalaenopsis orchids
Frontiers in Plant Science, 2018Co-Authors: Yu Chen Chuang, Wen Chieh Tsai, Yi Chu Hung, Masaru Ohmetakagi, Wen Huei Chen, Nobutaka Mitsuda, Honghwa ChenAbstract:Phalaenopsis bellina is a scented orchid emitting large amount of monoterpenes. GERANYL DIPHOSPHATE SYNTHASE (PbGDPS) is the key enzyme for monoterpene biosynthesis, and shows concomitant expression with the emission of monoterpenes during flower development in P. bellina. Here, we identified a dual repeat cis-element in the GDPS promoter that is critical for monoterpene biosynthesis in Phalaenopsis orchids. A strong correlation between the dual repeat and the monoterpene production was revealed by examination of the GDPS promoter fragments over 12 Phalaenopsis species. Serial-deletion of the 2-kb GDPS promoter fragments demonstrated that the integrity of the dual repeat was crucial for its promoter activities. By screening the Arabidopsis transcription factors (TFs) cDNA library using yeast one-hybrid assay, AtbZIP18, a member of group I of bZIP TFs, was identified to be able to bind the dual repeat. We then identified PbbZIP4 in the transcriptome of P. bellina, showing 83% identity in the DNA binding region with that of AtbZIP18, and the expression level of PbbZIP4 was higher in the scented orchids. In addition, PbbZIP4 transactivated the GDPS promoter fragment containing the dual repeat in dual luciferase assay. Furthermore, transient ectopic expression of PbbZIP4 induced a 10-fold production of monoterpenoids in the scentless orchid. In conclusion, these results indicate that the dual repeat is a real TF-bound cis-element significant for GDPS gene expression, and thus subsequent monoterpene biosynthesis in the scented Phalaenopsis orchids.
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ploidy doubling by in vitro culture of excised protocorms or protocorm like bodies in Phalaenopsis species
Plant Cell Tissue and Organ Culture, 2009Co-Authors: Wen Huei Chen, Ching Yan Tang, Yulin KaoAbstract:Endopolyploidy was observed in the protocorms of diploid Phalaenopsis aphrodite subsp. formosana with ploidy doubling achieved by in vitro regeneration of excised protocorms, or protocorm-like bodies (PLBs). Thirty-four per cent of the PLBs regenerated from the first cycle of sectioned protocorms were found to be polyploids with ploidy doubled once or twice as determined by flow-cytometry. The frequency of ploidy doubling increased as the sectioning cycles increased and was highest in diploid followed by the triploid and tetraploid. Regeneration of the endopolyploid cells in the tissue of the protocorms or PLBs is proposed as the source of the development of ploidy doubled plantlets. The frequency of ploidy doubling was similar in seven other Phalaenopsis species, although the rate of increase within cycles was genotype specific. In two species, a comparison of five parameters between 5-month-old diploid and tetraploid potted plants showed only the stomata density differed significantly. The flowers of the tetraploid plant were larger and heavier than those of the diploids. This ploidy doubling method is a simple and effective means to produce large number of polyploid Phalaenopsis species plants as well as their hybrids. The method will be beneficial to orchid breeding programs especially for the interspecific hybridization between varieties having different chromosome sizes and ploidy levels.
Ching-chung Chen - One of the best experts on this subject based on the ideXlab platform.
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Complete nucleotide sequence of capsicum chlorosis virus isolated from Phalaenopsis orchid and the prediction of the unexplored genetic information of tospoviruses
Archives of Virology, 2011Co-Authors: You-xiu Zheng, Ching-chung ChenAbstract:Phalaenopsis orchids are popular ornamentals all over the world. A tospovirus, capsicum chlorosis virus (CaCV-Ph) had been identified as the cause of chlorotic ringspots on leaves of Phalaenopsis orchids in Taiwan. The tripartite genome of CaCV-Ph was found to contain 3608, 4848 and 8916 nt of S, M and L RNAs, respectively. Phylogenetic analysis of the nucleocapsid (N) protein confirmed that CaCV-Ph is a member of the watermelon silver mottle virus (WSMoV) serogroup in the genus Tospovirus . Based on the relations among the nonstructural protein (NSs), glycoprotein (GnGc), thrips genera, host and geographical distribution, tospoviruses and thrips could be classified into two major types: WSMoV- Thrips -Asian and Tomato spotted wilt virus (TSWV)- Frankliniella -EuroAmerican. The proline (P_459) of all tospoviral Gn proteins was indispensable for thrips transmission, but the RGD motif, which is maintained by only six tospoviruses, may not be required for thrips transmission. An RdRp catalytic domain found in the conserved region of the L protein may recognize the typically conserved sequences on the 5’ and 3’ terminal regions (5’ AGAGCAAU 3’).
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Odontoglossum ringspot virus causing flower crinkle in Phalaenopsis hybrids
European Journal of Plant Pathology, 2010Co-Authors: You-xiu Zheng, Bing-nan Shen, Ching-chung ChenAbstract:A new disorder exhibiting flower crinkle on Phalaenopsis orchids bearing white flowers has been observed in Taiwan, China and Japan for several years. This disorder decreased the flower longevity and was considered as a physiological syndrome. The objective of this study was to identify and characterize the real causal agent of this new Phalaenopsis disorder. Five plants of Phalaenopsis hybrids “V3” (Phal. Yukimai × Phal. Taisuco Kochdian) with flower crinkle symptoms were collected and tested by enzyme-linked immunosorbent assay with antisera against 18 viruses. The extract of leaves and flowers from one diseased plant (96-Ph-16) reacted positively only to antiserum against Odontoglossum ringspot virus (ORSV), while those from the other four plants (96-Ph-7, 96-Ph-17, 96-Ph-18 and 96-Ph-19) reacted positively to the antisera against ORSV and Cymbidium mosaic virus (CymMV). Five ORSV isolates, one each from flowers of those five diseased Phalaenopsis orchids, were established in Chenopodium quinoa . A CymMV culture was isolated from the flowers of one of the ORSV/CymMV mix-infected Phalaenopsis orchids (96-Ph-19). To determine the causal agent of the flower crinkle disease, healthy Phalaenopsis seedlings were singly or doubly inoculated with the isolated ORSV and/or CymMV. Results of back inoculation indicated that ORSV is the sole causal agent of the crinkle symptom on petals of Phalaenopsis orchid. The CP gene of the ORSV isolates from this study shared 97.3–100% nucleotide identity and 96.2–100% amino acid identity with those of 41 ORSV isolates available in GenBank. This is the first report demonstrating ORSV as the sole virus causing flower crinkle disease on Phalaenopsis orchids.
Mahfut Mahfut - One of the best experts on this subject based on the ideXlab platform.
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Ceratorhiza Induction towards ORSV Infection on Viability of Phalaenopsis and Dendrobium
'LPPM Universitas Khairun', 2021Co-Authors: Sari, Valentina Dwi Anggita, Mahfut Mahfut, Wahyuningsih Sri, Handayani, Tundjung TripeniAbstract:Orchids have a high level of biodiversity, such as Dendrobium and Phalaenopsis, which are epiphytes. Dendrobium can adapt to the condition of where it lives while Phalaenopsis can grow in highlands and depends on sunlight and humidity. Virus infection has become one of the obstacles in cultivating Dendrobium and Phalaenopsis. Efforts to increase fitness and control in Dendrobium and Phalaenopsis cultivation can be done by inducing the plant’s fitness using a mycorrhiza, such as Ceratorhiza. A mycorrhiza is a form of mutualism between fungi and the plant’s root. This research is aimed to give information related to the utilization of Cerathoriza for inducing orchids to suppress Odontoglossum ringspot virus (ORSV) infection, giving it better growth. The research was done in February-March 2021 at Botany Laboratory University of Lampung. A completely randomized factorial design was used on two factors, kind of orchid and mycorrhiza treatment (M), virus (V), and mycorrhiza virus (MV). Variables examined in this research are the amount of living and dead roots and leaves. Data obtained is homogenized using Levene’s test and continued by ANOVA with the significance level of 5% and further testing using Tukey’s test with the significance level of 5%. From this research, it is known that interaction between Phalaenopsis and Dendrobium exists during virus and mycorrhiza administration. It is concluded that Phalaenopsis anabilis is more vulnerable than Dendrobium discolor
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Various ORSV infection symptoms on ceratorhiza induction in orchid plants
'Lembaga Penelitian dan Pengabdian kepada Masyarakat Universitas Lampung', 2021Co-Authors: Panjaitan, Mitha Valentina Treesya, Mahfut Mahfut, Handayani, Tundjung Tripeni, Wahyuningsih SriAbstract:Several types of orchids that are in great demand and cultivated by various groups are Phalaenopsis sp. and Dendrobium sp. Mycorrhizae are needed by orchid plants to complete their life cycle. This is because mycorrhizae are capable of symbiosis with the root system of the host plant so that the plant's capacity to absorb elements will increase. This study aims to determine the extent to which Ceratorhiza induction on Phalaenopsis amabilis and Dendrobium discolour orchids can suppress the symptoms of Odontoglossum ringspot virus (ORSV) infection. The method used in this study was a factorial completely randomized design (CRD) with 2 factors and 4 replications. The first factor is the type of orchid used, namely Phalaenopsis amabilis (A1) and Dendrobium discolour (A2), while the second factor is the type of administration of mycorrhizae, viruses, and mycorrhizal viruses. Observation of disease symptoms is done by looking at the symptoms that appear on leaves that have been infected with ORSV. The results showed that the symptoms that appeared in Phalaenopsis amabilis were in the form of mosaic, necrosis, leaf malformation, while in Dendrobium discolour were mosaic and necrotic. Ceratorhiza was not able to suppress the symptoms of disease infection because based on the results obtained; the treatment on orchids inoculated with mycorrhizae and viruses showed more severe symptoms compared to the treatment only inoculated with viruses increased response and Dendrobium discolours showed a decreased response in leaf length, width, and number. Meanwhile, in the root organ, Dendrobium discolour showed an increased response and Phalaenopsis amabilis showed a decreased response in root length and number of roots
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Variation of resistance response of orchids induced by Rhizoctonia against the infection of Odontoglossum ringspot virus (ORSV) based on percentage of disease development
'Lembaga Penelitian dan Pengabdian kepada Masyarakat Universitas Lampung', 2021Co-Authors: Izzati, Fania Nur, Mahfut Mahfut, Ernawiati Eti, Wahyuningsih SriAbstract:Orchid is one of the largest groups of flowering plants that are in great demand by society. Besides, the various flowers with a long vase life, the price of orchids is also stable and affordable. Phalaenopsis and Dendrobium are the most popular types of orchids. To anticipate this, orchid cultivation needs to be improved. However, orchid cultivation is not always accompanied by the effort of disease prevention, particularly those caused by viruses. The virus that most infects orchids is the Odontoglossum ringspot virus (ORSV). Symptoms that appear on the leaves are chlorosis, mosaic with a pattern of lines, circles, and necrotic rings/rings. Control of viral infections can utilize organisms that are able to induce resistance, namely mycorrhizae. Mycorrhizae will penetrate into the root cortex tissue and provide nutrients for the orchid. Mycorrhizae that are quite often found are Rhizoctonia and are applied by induction to orchids. Induction was undertaken by planting orchids in Rhizoctonia inoculum until anatomically a peloton structure (solid coils) was formed on the orchid roots. The aim of this study was to study the response of orchid plants in the form of percentage incidence and intensity of the disease and the level of plant resistance to ORSV infection in Phalaenopsis amabilis and Dendrobium discolour which had been induced by Rhizoctonia. This research was conducted at the Laboratory of Botany, Biology, FMIPA, Universitas Lampung, using a factorial completely randomized design (CRD). Factor 1 was the type of orchid and factor 2 was mycorrhizal induction, virus inoculation, and a combination of both with 4 replications. The finding indicated that Phalaenopsis amabilis was more susceptible to ORSV infection than Dendrobium discolour based on a higher incidence and intensity of the disease. Mycorrhizal induction and virus inoculation (MAV) in Dendrobium discolour showed an increase of resistance response compared to Phalaenopsis amabilis
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Induction of Rhizoctonia resistance to odontoglossum ringspot virus (ORSV) infection on orchid leaf organ viability
'Lembaga Penelitian dan Pengabdian kepada Masyarakat Universitas Lampung', 2021Co-Authors: Tohari, Eka Nuraini, Mahfut Mahfut, Handayani, Tundjung Tripeni, Wahyuningsih SriAbstract:Orchid plants are in great demand by the public as ornamental plants that have economic value and the beauty of attractive flower shapes and colors, especially Phalaenopsis and Dendrobium. Related to this, conservation efforts are needed through cultivation. However, these efforts are often hampered by viral infections. One of the viruses that trigger inhibition of orchid cultivation that often appears is ORSV. Control efforts that are relatively safe and efficient can utilize mycorrhizae in orchids which play a role in increasing the absorption of plant nutrients. The use of Phalaenopsis and Dendrobium with the induction of Rhizoctonia sp. against ORSV is a very important alternative biofertilizer and biocontrol agent. The objectives of this study were 1) to determine the effectiveness of orchids induced by Rhizoctonia resistance, 2) to determine the viability of orchid leaf organs to ORSV infection, 3) to determine the viability of orchid root organs resulting from Rhizoctonia resistance to ORSV infection. This study was conducted in January 2021 – March 2021 at the Botanical 2 Laboratory, Department of Biology, Faculty of Mathematics and Natural Sciences, University of Lampung. This study employed a completely randomized factorial with 6 treatments and 4 replications. The variable observed in this study was the effectiveness of mycorrhizae, leaf and root length, number of leaves and roots, and leaf width. The data obtained were homogenized using Levene's test and then analyzed by ANOVA and Tukey's further test at the 5% level. The results showed that the effectiveness of mycorrhizae occurred on the 3rd day of incubation. In leaf organs, Phalaenopsis amabilis showed an increased response and Dendrobium discolour showed a decreased response in leaf length, width, and number. Meanwhile, in the root organ, Dendrobium discolour showed an increased response and Phalaenopsis amabilis showed a decreased response in root length and number of roots.
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Variation Symptoms and Resistance Response of Different Types on Orchids (Orchidaceae) Against Odontoglossum ringspot virus (ORSV) Infection
'Lembaga Penelitian dan Pengabdian kepada Masyarakat Universitas Lampung', 2020Co-Authors: Mahfut Mahfut, Daryono, Budi SetiadiAbstract:Odontoglossum ringspot virus (ORSV) is one of the viruses that are reported to infect the most orchids in Indonesia. Viral infections cause economic losses. This research was conducted to study the introduction of symptoms of infection that appear as the initial stages of disease control. The source of the virus is orchid leaves Phalaenopsis amabilis positively infected by ORSV collected from Magelang. Virus isolation was carried out by mechanical inoculation to indicator plants (Chenopodium amaranticolor and Nicotiana tabaccum). Furthermore, the results of inoculation were multiplied using various types of orchids (Dendrobium, Cymbidium, Chattleya, Phalaenopsis, Spatthoglotis, Liparis, and Pecteilis). Virus detection on orchid tissue was done trough the DAS-ELISA and observation of virus structure using TEM. The results showed that Pectelis sussanae (L.) Raf. was the only orchid plant that was immune against ORSV approached. Dendrobium sp., Cymbidium sp., Chattleya sp., Phalaenopsis sp., and Spathoglotis sp. showed a response of susceptible of ORSV severe infection symptoms, while Liparis sp. were resistant. The variation of resistance from several orchid plant against infection ORSV showed severe symptoms with the incubation periode was seen earlier. The observation of virus structure using TEM showed rigid road shape particle, 300 x 18 nm in size, which is general characteristic of Tobamovirus. This indicate an infection of ORSV is a dangerous disease and require serious contro
