Agrobacterium tumefaciens telah digunakan secara luas pada penelitian molekuler tanaman dan pemul... more Agrobacterium tumefaciens telah digunakan secara luas pada penelitian molekuler tanaman dan pemuliaan tanaman sejak tahun 1983 untuk kepentingan rekayasa genetika. Umumnya A.tumefaciens diinfeksi pada diskus daun sedangkan menginjeksi A.tumefaciens ke jaringan daun dapat dijadikan alternatif, metode ini disebut Agroinfiltrasi. Agroinfiltrasi lebih cepat dan mudah dibandingkan teknis diskus daun karena dapat mengukur ekspresi transgene sejak 2-4 hari setelah infiltrasi. Sedangkan metode diskus daun memerlukan waktu 2-4 bulan untuk analisa ekspresi transgene. Pada penelitian ini, Agroinfiltrasi daun dilakukan pada Sindoro salah satu jenis tembakau Temanggung. Variabel agroinfiltrasi adalah densitas inokulum A.tumefaciens C58C1(pMp90, pEGAD) dan waktu ko-kultivasi setelah infiltrasi. Kuantifikasi Green Fluorescent Protein (GFP) ekstrak daun Sindoro dibandingkan kontrol negatif dapat mencapai 123% pada OD600=0.06 saat 24 jam setelah infiltrasi. Sebagian agroinfiltrasi daun Sindoro juga diregenerasi pada media Murashige dan Skoog (MS) mengandung BASTA 5 ppm sebagai agen seleksi. Tunas transforman muncul pada hari ke-47 setelah inokulasi di media MS seleksi transforman.
This study aimed to clone thexynB gene from Bacillussubtilissubsp. spizizeniiW23,encodinga xylan ... more This study aimed to clone thexynB gene from Bacillussubtilissubsp. spizizeniiW23,encodinga xylan l,4-beta-xylosidase to pMMB67EH plasmid which then be used to transformed EscherichiacoliDH-5a and Origami host cells. The xynBgenewas amplifiedby polymerasechain reaction (PCR) technique using a pair of primers flanking the gene sequence,and chromosomal DNA of the W23 strain as a template. Analyses of the recombinant plasmid were done by restriction analyses, and PCR detection. The result showed that the xynB has been cloned on pMMB67EH vector. The recombinant plasmid contained the xynB gene which was confirmed by restriction analyses and by PCR detection using primers pair's specific for the xynB gene and for the vector. The xylanaseactivity of xynB gene in E.coliDH-5a and Origami host cells was assayedon luria-Bertani-xylan plate qualitatively with addition of isopropyl B-D-thio-galactoside (IPTG) as an inducer.Uponsprayingwith Congored, the cells bearing the pMMB-xynB recombinant plasmid showed a xylan-degrading activity by the appearance ofclear zanearound the colonies whilethe transformant bearing an empty plasmid showed no clear zone. It could be concluded that the cloning Arocess was succeeded.
Mata kuliah Teknik Analisa DNA mengajarkan kepada mahasiswa mengenai teknik-teknik analisa Biolog... more Mata kuliah Teknik Analisa DNA mengajarkan kepada mahasiswa mengenai teknik-teknik analisa Biologi Molekuler, khususnya DNA. Namun, rancangan perangkat kurikulum dan metode pengajaran yang sebelumnya diterapkan masih menitikberatkan pada prosedur, sehingga minat dan motivasi belajar mahasiswa menjadi rendah. Inovasi pembelajaran ini bertujuan untuk menata ulang perangkat kurikulum berdasarkan metode pembelajaran kontekstual serta merancang metode dan media pembelajaran yang mengacu pada Student Centered Learning (SCL) yang diharapkan dapat meningkatkan minat dan motivasi belajar mahasiswa. Pembelajaran kontekstual dilakukan melalui pemberian case study dan proyek yang mengaplikasikan teknik analisa DNA untuk mencari solusi atas suatu permasalahan. Hasil implementasi inovasi pembelajaran ini menunjukkan rerata capaian ketuntasan belajar sebesar 61,28% dan peningkatan motivasi belajar mandiri mahasiswa sebesar 51,25%. Selain itu, juga terjadi peningkatan kepuasan mahasiswa terhadap ki...
La reponse hypersensible ou hr, caracterisee par la mort des cellules vegetales au point d'in... more La reponse hypersensible ou hr, caracterisee par la mort des cellules vegetales au point d'infection, et conduisant au confinement de l'agent pathogene, est generalement associee a la resistance des plantes aux agents pathogenes. Cette reponse fait appel a un programme genetique de mort cellulaire, et si les evenements precoces conduisant au declenchement de la hr sont de mieux en mieux connus, l'execution du programme de mort cellulaire demeure en revanche une enigme. Le travail presente dans ce manuscrit est focalise sur un acteur potentiel de ce programme chez arabidopsis thaliana, le gene atmyb30. Ce gene avait ete isole au laboratoire sur la base de son expression, induite specifiquement lors d'une interaction incompatible avec la bacterie pathogene xanthomonas campestris pv. Campestris, et appartient a la famille des facteurs transcriptionnels de type myb. Une etude precise de son expression a montre qu'elle est specifique, precoce et transitoire lors de la...
An attack of plants by pathogens or treatment with certain resistance-inducing compounds can lead... more An attack of plants by pathogens or treatment with certain resistance-inducing compounds can lead to the establishment of a unique primed state of defense. Primed plants show enhanced defense reactions upon further challenge with biotic or abiotic stress. Here, we report that the primed state in Arabidopsis (Arabidopsis thaliana) is still functional in the next generation without additional treatment. We compared the reactions of Arabidopsis plants that had been either primed with β-amino-butyric acid (BABA) or with an avirulent isolate of the bacteria Pseudomonas syringae pv tomato (PstavrRpt2). The descendants of primed plants showed a faster and higher accumulation of transcripts of defense-related genes in the salicylic acid signaling pathway and enhanced disease resistance upon challenge inoculation with a virulent isolate of P. syringae. In addition, the progeny of primed plants was also more resistant against the oomycete pathogen Hyaloperonospora arabidopsidis. When transgenerationally primed plants were subjected to an additional priming treatment, their descendants displayed an even stronger primed phenotype, suggesting that plants can inherit a sensitization for the priming phenomenon. Interestingly, this primed to be primed phenotype was much reduced in the Arabidopsis β-amino-butyric acid priming mutant ibs1 (induced BABA sterility1). Our results demonstrate that the primed state of plants is transferred to their progeny and confers improved protection from pathogen attack as compared to the descendants of unprimed plants.
Agrobacterium tumefaciens telah digunakan secara luas pada penelitian molekuler tanaman dan pemul... more Agrobacterium tumefaciens telah digunakan secara luas pada penelitian molekuler tanaman dan pemuliaan tanaman sejak tahun 1983 untuk kepentingan rekayasa genetika. Umumnya A.tumefaciens diinfeksi pada diskus daun sedangkan menginjeksi A.tumefaciens ke jaringan daun dapat dijadikan alternatif, metode ini disebut Agroinfiltrasi. Agroinfiltrasi lebih cepat dan mudah dibandingkan teknis diskus daun karena dapat mengukur ekspresi transgene sejak 2-4 hari setelah infiltrasi. Sedangkan metode diskus daun memerlukan waktu 2-4 bulan untuk analisa ekspresi transgene. Pada penelitian ini, Agroinfiltrasi daun dilakukan pada Sindoro salah satu jenis tembakau Temanggung. Variabel agroinfiltrasi adalah densitas inokulum A.tumefaciens C58C1(pMp90, pEGAD) dan waktu ko-kultivasi setelah infiltrasi. Kuantifikasi Green Fluorescent Protein (GFP) ekstrak daun Sindoro dibandingkan kontrol negatif dapat mencapai 123% pada OD600=0.06 saat 24 jam setelah infiltrasi. Sebagian agroinfiltrasi daun Sindoro juga diregenerasi pada media Murashige dan Skoog (MS) mengandung BASTA 5 ppm sebagai agen seleksi. Tunas transforman muncul pada hari ke-47 setelah inokulasi di media MS seleksi transforman.
This study aimed to clone thexynB gene from Bacillussubtilissubsp. spizizeniiW23,encodinga xylan ... more This study aimed to clone thexynB gene from Bacillussubtilissubsp. spizizeniiW23,encodinga xylan l,4-beta-xylosidase to pMMB67EH plasmid which then be used to transformed EscherichiacoliDH-5a and Origami host cells. The xynBgenewas amplifiedby polymerasechain reaction (PCR) technique using a pair of primers flanking the gene sequence,and chromosomal DNA of the W23 strain as a template. Analyses of the recombinant plasmid were done by restriction analyses, and PCR detection. The result showed that the xynB has been cloned on pMMB67EH vector. The recombinant plasmid contained the xynB gene which was confirmed by restriction analyses and by PCR detection using primers pair's specific for the xynB gene and for the vector. The xylanaseactivity of xynB gene in E.coliDH-5a and Origami host cells was assayedon luria-Bertani-xylan plate qualitatively with addition of isopropyl B-D-thio-galactoside (IPTG) as an inducer.Uponsprayingwith Congored, the cells bearing the pMMB-xynB recombinant plasmid showed a xylan-degrading activity by the appearance ofclear zanearound the colonies whilethe transformant bearing an empty plasmid showed no clear zone. It could be concluded that the cloning Arocess was succeeded.
Mata kuliah Teknik Analisa DNA mengajarkan kepada mahasiswa mengenai teknik-teknik analisa Biolog... more Mata kuliah Teknik Analisa DNA mengajarkan kepada mahasiswa mengenai teknik-teknik analisa Biologi Molekuler, khususnya DNA. Namun, rancangan perangkat kurikulum dan metode pengajaran yang sebelumnya diterapkan masih menitikberatkan pada prosedur, sehingga minat dan motivasi belajar mahasiswa menjadi rendah. Inovasi pembelajaran ini bertujuan untuk menata ulang perangkat kurikulum berdasarkan metode pembelajaran kontekstual serta merancang metode dan media pembelajaran yang mengacu pada Student Centered Learning (SCL) yang diharapkan dapat meningkatkan minat dan motivasi belajar mahasiswa. Pembelajaran kontekstual dilakukan melalui pemberian case study dan proyek yang mengaplikasikan teknik analisa DNA untuk mencari solusi atas suatu permasalahan. Hasil implementasi inovasi pembelajaran ini menunjukkan rerata capaian ketuntasan belajar sebesar 61,28% dan peningkatan motivasi belajar mandiri mahasiswa sebesar 51,25%. Selain itu, juga terjadi peningkatan kepuasan mahasiswa terhadap ki...
La reponse hypersensible ou hr, caracterisee par la mort des cellules vegetales au point d'in... more La reponse hypersensible ou hr, caracterisee par la mort des cellules vegetales au point d'infection, et conduisant au confinement de l'agent pathogene, est generalement associee a la resistance des plantes aux agents pathogenes. Cette reponse fait appel a un programme genetique de mort cellulaire, et si les evenements precoces conduisant au declenchement de la hr sont de mieux en mieux connus, l'execution du programme de mort cellulaire demeure en revanche une enigme. Le travail presente dans ce manuscrit est focalise sur un acteur potentiel de ce programme chez arabidopsis thaliana, le gene atmyb30. Ce gene avait ete isole au laboratoire sur la base de son expression, induite specifiquement lors d'une interaction incompatible avec la bacterie pathogene xanthomonas campestris pv. Campestris, et appartient a la famille des facteurs transcriptionnels de type myb. Une etude precise de son expression a montre qu'elle est specifique, precoce et transitoire lors de la...
An attack of plants by pathogens or treatment with certain resistance-inducing compounds can lead... more An attack of plants by pathogens or treatment with certain resistance-inducing compounds can lead to the establishment of a unique primed state of defense. Primed plants show enhanced defense reactions upon further challenge with biotic or abiotic stress. Here, we report that the primed state in Arabidopsis (Arabidopsis thaliana) is still functional in the next generation without additional treatment. We compared the reactions of Arabidopsis plants that had been either primed with β-amino-butyric acid (BABA) or with an avirulent isolate of the bacteria Pseudomonas syringae pv tomato (PstavrRpt2). The descendants of primed plants showed a faster and higher accumulation of transcripts of defense-related genes in the salicylic acid signaling pathway and enhanced disease resistance upon challenge inoculation with a virulent isolate of P. syringae. In addition, the progeny of primed plants was also more resistant against the oomycete pathogen Hyaloperonospora arabidopsidis. When transgenerationally primed plants were subjected to an additional priming treatment, their descendants displayed an even stronger primed phenotype, suggesting that plants can inherit a sensitization for the priming phenomenon. Interestingly, this primed to be primed phenotype was much reduced in the Arabidopsis β-amino-butyric acid priming mutant ibs1 (induced BABA sterility1). Our results demonstrate that the primed state of plants is transferred to their progeny and confers improved protection from pathogen attack as compared to the descendants of unprimed plants.
Uploads
Papers
Teaching Documents