International Cooperation

Practical Courses - Conference

4th course "The NIBB Laboratory Course and Workshops on Physcomitrella patens 2009"


Organizers	Andrew Cuming (Univ. Leeds) Yuji Hiwatashi (NIBB) Yasuko Kamisugi (Univ. Leeds) Minoru Kubo (ERATO, JST) Tetsuya Kurata (ERATO, JST) Takashi Murata (NIBB) Tomoaki Nishiyama (Kanazawa Univ.) Keiko Sakakibara (ERATO, JST) Yoshikatsu
Venue	National Institute for Basic Biology
Date	Jun. 29-Jul. 3, 2009
Link	Website　（http://www.nibb.ac.jp/course/past/4th/）
	Website　（http://www.nibb.ac.jp/course/past/4th/）
Poster

Ever since the first NIBB International Practical Course, held in 2007, NIBB has continued to provide young scientists from around the world with opportunities to practice cutting-edge skills and to spend valuable time learning from experts in the field. For twenty years prior to 2007 NIBB held Bioscience Training Courses to offer these same opportunities, however they were attended mostly by the students of universities and institutes in Japan. The International Practical Course program was initiated to bring these goals to the world stage and offer the opportunities available at the Bioscience Training Courses to future researchers without regard to nationality. The Fourth International Practical Course was held from June 29th to July 3rd, 2009, focusing on techniques used in the biology of the moss Physcomitrella patens, an important model organism in plant biology.

Seventeen students participated in the course, from Japan, Israel, Germany, Belgium, The Czech Republic, France, Sweden, Singapore, and India. They worked on learning new techniques and capitalized on the many chances to enjoy discussions with established researchers throughout the five days of the course.

The course provided basic techniques including cultivation, observation of developmental processes at the cellular level, gene targeting, bioimaging, and bioinformatics. The course was taught, with the help of laboratory technicians, by Drs. Andrew C. Cuming (Univ. Leeds), Yasuko Kamisugi (Univ. Leeds), Tomoaki Nishiyama (Kanazawa Univ.), Tetsuya Kurata (JST), Yoshikatsu Sato (JST), Yuji Hiwatashi (NIBB), Minoru Kubo (JST), Takashi Murata (NIBB), Mitsuyasu Hasebe (NIBB). In-addition many postdoctoral fellows working at NIBB introduced the research they are perusing using P. patens and shared detailed research procedures with the participants.

The laboratories used were provided by NIBB and equipped with the necessary instruments and tools as well as a good supply of material moss. We hope that the techniques and knowledge provided in this course will prove useful in the future work of all of the participants.

Program

Program
June 28, Sun
17:00	Get together

June 29, Mon
09:00	Welcome address (Kiyotaka Okada, Director General of NIBB)
09:05	Introduction of course and Physcomitrella patens	Hasebe and Cuming
09:35	Spore germination	Hiwatashi
	Culture and storage of protonemata and gametophores	Hiwatashi
10:15	Observation of protonemata and gametophores	Sato
	Analysis of protonema colony morphology	Sato
	Regeneration of protonemata from excised leaves-1	Sato
11:05	Induction of gametangia and sporophytes	Cuming and Kamisugi
	Sporangium collection and storage	Cuming and Kamisugi
	Observation of antheridia and archegonia	Cuming and Kamisugi
	Observation of sperms	Cuming and Kamisugi
12:05	Lunch
13:00	PEG-mediated transformation	Hiwatashi, Cuming and Kamisugi
17:00	Crosses of mutants defective in the sporophyte formation	Cuming and Kamisugi
18:00	Dinner	Kurata
20:00	Workshop 1: Next generation sequencer	Kurata

June 30, Tue
09:00	PEG-mediated transformation	Hiwatashi, Cuming and Kamisugi
13:00	Lunch
14:00	Genomic DNA extraction	Sakakibara
	Chromatin immunoprecipitation	Kurata
	RNA extraction	Kurata
	Green PCR	Kubo
15:00	DNA-gel-blot analysis	Sakakibara
	RNA interference	Kurata
	Artificial micro RNA	Kurata
	Gene deletion	Sakakibara
	Conditional deletion	Kubo and Sato
	Dominant repression	Kubo
	Overexpression	Kubo
	Inducible expression	Kubo
18:00	Dinner
20:00	Workshop 2: Targeting strategy	Cuming and Kamisugi

July 1, Wed
09:00	PEG-mediated transformation	Hiwatashi, Cuming and Kamisugi
11:00	How to use genome information	Nishiyama
12:00	Lunch
	Free time / Seimar by Dr. Nir Ohad and Dr. Amal J. Johnston

July 2, Thu
09:00	How to use genome information	Nishiyama
10:00	Analysis of protonema development and growth	Sato and Hiwatashi
11:00	Observation of microtubules with indirect immunofluorescence microscopy	Murata
	Light and electron microscopy of protonemata embedded with epoxy resin	Murata
12:00	Lunch
13:00	DAPI, Hoechst33342, and PI staining of protonemata	Murata
	Visualization of cell wall with calcoflur	Murata
14:00	Live imaging	Sato
	Cellular localization of fluorescent protein	Sato
	Regeneration of protonemata from excised leaves-2	Sato
16:00	GUS staining	Hiwatashi
17:00	How to use genome information	Nishiyama
18:00	Dinner
20:00	Workshop 3: Bioimaging	Sato, Murata, Cuming and Kamisugi
July 3, Fri
09:00	Measurement of DNA content with FACS	Sato
10:00	Agrobacteria-mediated moss transformation: Protonema and Agrobacteria Co-culture	Kurata
12:00	Lunch
13:00	Agrobacteria-mediated moss transformation: Protonema and Agrobacteria Co-culture	Kurata
14:00	Hormone treatments	Cuming and Kamisugi
15:00	Light response	Sato
	Gravitropic response	Sato
16:00	Closing discussion	All lecturers
17:00	Farewell party