김민지 방미라 이정민 조경국 2006 정호영

Title Authors

- Table of Contents - AbstractAbstract IntroductionIntroduction Materials and MethodsMaterials and Methods ResultsResults DiscussionDiscussion ConclusionConclusion

1. Abstract A protocol was established for cotton somatic embryogenesis and highly efficient plant regeneration. 20 Chinese and Australian commercialized cotton cultivars including CCRI 12, CCRI 19, and Simian No 3.

1. Abstract Simplifies cotton somatic embryogenesis From a multi-step to one- step culture process and shortens the culture cycle from 180 to days The application of plant genetic engineering on cotton genetic improvement.

2. Introduction Attempted to investigate the effect of different factors (genotypes, explants, plant growth regulators) Developed a simple procedure for highly efficient and rapid plant regeneration For current commercialized cotton cultivars

3. Material & Method Plant materials and the establishment of sterile seedlings –Chinese commercialized cotton cultivars CCRI 12, CCRI-13, CCRI 16, CCRI 17, CCRI 19, CCRI 24, CCRI 27, CCRI 29, CCRI 30, Simian No 3, Simian No 4,Lumian 1, Lumian 6, Jinmian 7, Jinmian 12, Jihe 321, Yumian 8 –The Australia commercialized cotton Cultivars Siokral 1-3 and Siokral 1-4; and the elite regenerable cotton cultivars Coker 201 and Coker 312

3. Material & Method Plant materials and the establishment of sterile seedlings ① Kernels were removed from the seeds. ② Mature kernels were chosen and were surface- sterilized. ③ The kernels were then cultured 0.7% agar- solidified medium, pH 5.8. ④ The kernels were cultured at 28°C ± 2°C under 16 h light photoperiod conditions with a light intensity of approximately 24 PPFD.

3. Material & Method Induction of callus –Hypocotyl sections –Cotyledon pieces –Root segments ① 7-day-old sterile seedlings were cultured for inducing callus..

3. Material & Method Selection of high frequency embryogenic cell lines ① After days of culture, embryogenic callus was chosen and transferred. a.MSB medium : different hormones b.MSB medium without hormon es ② After 28 days of subculture, embryogenic callus with a high frequency of embryogenesis was chosen for the next subculture. ③ Subsequently, embryogenic callus was subcultured every 28 days. a.MSB medium : 0.1 mg/L ZT and 2 g/L activated charcoal.

3. Material & Method Differentiation of somatic embryos and plant regeneration High frequency embryogenic cell lines were chosen and transferred onto embryo differentiation medium. –MSB medium mg/L ZT Mature embryos were chosen and transferred onto embryo germination medium after 30 days. –MSB or ZH medium mg/L ZT and 2 g/L activated charcoal.

3. Material & Method Experimental setup and statistical analysis ① Repeat 3-4 ② Standard statistical software (SPSS, Chicago, Illinois, USA) ③ The Least Significant Difference (LSD)

4. Results

5. Discussion (Past) Difficult to induce somatic embryogenesis and plant regeneration in cotton. Have been used to obtain genetically modified cotton (Agrobacterium, particle bombardment) Three obstacles - Long periods of tissue culture - Frequency is very low - Genotype dependent.

5. Discussion (Past) Commercialized cultivars, the frequency of plant regeneration has been low - Regeneration procedure has required multiple subcultures that often produced unwanted somaclonal variation. - Genotype-dependent plant regeneration response. Transgenic cotton plants must be crossed with commercialized cotton cultivars

5. Discussion Develop a simple procedure for cotton plant regeneration via somatic embryogenesis ZT and cytokinin can induce somatic embryogenic callus

5. Discussion

6. Conclusion Plant regeneration via somatic embryogenesis in cotton efficient and rapid

Thank you Thank you Q&A