Development of an Efficient Plant Regeneration System from Apomictic and Sexual Seeds of Guinea Grass (Panicum maximum) | Chapter 8 | Research Perspective on Biological Science Vol. 1

Guinea grass (Panicum maximum) is an important tropical forage crop. It is represented in nature by both of the tetraploid biotypes that can reproduce through apomixis, and the sexual one. If this trait can be applied, it is expected to be an epoch-making breeding method, such as significantly reducing the cost of seed production by fixing first-generation hybrids, significantly shortening the breeding period by fixing promising strains in the middle of breeding, and converting vegetatively propagated plants to seed-propagated plants. The authors have captured a candidate gene, ASG-1, from apomictic guinea grass.　For analysis of apomixis genes, as the first step, an efficient and simple plant regeneration system has been established using callus from apomictic and sexual seeds of guinea grass (Panicum maximum). This study was conducted at the Miyakonojo Campus of Minami Kyushu University in Miyakonojo City, Japan, located at 31 degrees north latitude. The best basic medium for callus formation from mature seeds of guinea grass was selected, and then, the best combinations of growth regulators on different media were selected for plant regeneration by indirect organogenesis. Guinea grass accessions of sexual and apomict seeds were used for culture. Seeds sterilized were cultured in Murashige and Skoog (1962) medium (MS) and in that proposed by Chu et al. (1975) (N6D) for callus formation. The best medium and the effects of L-proline and growth regulators’ type and concentrations on callus formation and plant regeneration in 3 accessions were examined, respectively. After the plantlets rooted in MS hormone-free medium, the complete plants were planted in pots for hardening. As a result, N6D medium has given better rates for callus formation, that is, 97.1% in sexual N68/96-8-o-5, and 91.7% and 84.6% in apomict N68/96-8-o-11 and ‘Natsukaze’, respectively. MS medium with 0.2 mg/l of Kinetin has given the best rate of plant regeneration among the 4 kinds of media. For each material, the best results were obtained on MS with 0.2 mg/l of Kinetin for N68/96-8-o-5, and MS with 2.0 mg/l of L-proline and 0.2 mg/l of Kinetin for ‘Natsukaze’. After hardening of the regenerated plants in soil, 100% of survival rates were obtained, and showed normal growth compared with the plants derived from seeds. As described above, it was established, as the first case, an efficient and simple plant regeneration system by using callus from apomictic and sexual seeds of guinea grass for analysis of apomixis genes, consisting of analysis of the best media, L-proline usages and phytohormone combinations for callus formation, plant regeneration and hardening in different stages, respectively. This finding provides important information not only for sexual but also for apomictic plants in constructing a plant regeneration system from calli derived from mature seeds.

 

Author (s) Details

 

Chen Lanzhuang
Faculty of Environmental and Horticultural Science, Minami Kyushu University, 3764-1, Tatenocho, Miyakonojo City, Miyazaki, 885-0035, Japan.

Nishimura Yoshiko
Faculty of Environmental and Horticultural Science, Minami Kyushu University, 3764-1, Tatenocho, Miyakonojo City, Miyazaki, 885-0035, Japan

 

Umeki Kazuma
Faculty of Environmental and Horticultural Science, Minami Kyushu University, 3764-1, Tatenocho, Miyakonojo City, Miyazaki, 885-0035, Japan.

 

Zhang Jun
Qinghai Academy of Animal Science and Veterinary Medicine, Xining, Qinghai, 810016, China.

 

Xu Chengti
Qinghai Academy of Animal Science and Veterinary Medicine, Xining, Qinghai, 810016, China.

 

Please see the book here:- https://doi.org/10.9734/bpi/rpbs/v1/4894

A New Technology for Isolation and Manipulation of Single Protoplasts of Apomictic Reproductive Cells in Panicum maximum | Chapter 7 | Microbiology and Biotechnology Research: An Overview Vol. 1

Apomixis is a mode of reproduction in which only the mother's genes are transmitted from generation to generation. If this trait can be exploited, it can be expected to have an economic effect that exceeds that of the "Green Revolution".Apomixis is divided into three types from a breeding point of view, that is, adventitious embryogenesis, apospory and diplospory. The latter two belong to the gametophyte type. The reproductive mode of the gametophyte type of apomixis is basically divided into two parts. The first is the formation of an embryo sac at the 2n level; the second is that the egg cell of the embryo sac automatically develops into an embryo without fertilization. Guinea grass (Panicum maximum Jacq.) belonging to apospory was used and this study was carried out at Minami Kyushu University located in 31-degree north latitude of Japan. To do the molecular analysis of the mechanism of aposporous embryo sac initial cell (AIC) appearance, as the first step, we attempted to establish the system of isolating and manipulating single cells containing AIC using different methods in P. maximum. At first, single protoplasts were isolated from the ovaries staged in different developmental stages using different pre-treatments and different concentrations of enzyme solutions on ovaries; Based on this, the single protoplasts containing AIC were manipulated with ultra particle electronic syringe picopipet (UPESP) machine set onto microscope. The ovary of facultatively apomictic guinea grass (Panicum maximum) was used as a material in this study for single protoplast isolation. The ovaries were classified and collected from young buds and flowers staged in different developmental stages, according to the colors of stigma and the ovary length. Then, the ovaries were pre-treated with a needle into different shapes, and treated with different kinds of enzyme solution with different concentrations of mannitol to increase the efficiency of protoplast isolation. In final, the single protoplasts containing AIC isolated from different stages of ovaries were manipulated by handle control. The ovaries in different stages before AIC appearance, AIC appearance, and AIC-derived embryo sac formation were collected successfully and respectively, indicating that the stigma colors and length of ovaries are proportionate to stages of ovary maturity. And single protoplasts containing AIC were isolated from the ovaries staged in white to yellow color. 2) The ovaries were flooded in an enzyme solution and were pre-treated with a needle in 4 types, that is, (1) Cut in the micropylar end; (2) Cut in chalazal end; (3) Cut in the middle part; (4) after 1hr of (3) treatment, cut in micropylar end. As a result, the efficiency of protoplast isolation of (3) and (4) was 1-2 hrs shorter than that of (1) and (2). 12%, 11%, 10% and 9% were proper enzyme concentrations for obtaining perfect shingle protoplasts from the ovaries with white, yellow, peach and purple colors, respectively. 3) The single protoplasts containing AIC were collected and manipulated with UPESP in the performance of controlled aspiration and spit. This topic, in molecular analysis of the mechanism of AIC appearance of view, is relevant and promising, the key points were to isolate AIC single protoplasts from apomictic guinea grass using different methods, and then to establish the method of controlling a single protoplast using UPESP machine. The findings contribute to broader research on isolating and manipulation of reproductive cells, offering a useful tool for molecular analysis of AIC, and providing important information for clarification of apomixis reproductive mode which has potential implications for breeding strategies aimed at fixing F₁ hybrid crops.

 

Author (s) Details

Lanzhuang Che
Laboratory of Plant Biotechnology, Faculty of Environmental and Horticultural Science, Minami Kyushu University, 3764-1, Tatenocho, Miyakonojo city, Miyazaki 885-0035, Japan.

 

Yoshiko Nishimura
Laboratory of Plant Biotechnology, Faculty of Environmental and Horticultural Science, Minami Kyushu University, 3764-1, Tatenocho, Miyakonojo city, Miyazaki 885-0035, Japan.

 

Chenti Xu
Qinghai Academy of Animal Science and Veterinary Medicine, Xining, Qinghai 810016, China.

 

 

Please see the book here:- https://doi.org/10.9734/bpi/mbrao/v1/4895

Flower to Fruit Development in Coscinium fenestratum (Gaertn.) Colebr | Chapter 10 | Advanced Research in Biological Science Vol. 2

 This branch highlights the flower to crop development in Coscinium fenestratum (Gaertn.) Colebr., as an extensively secondhand slow growing class often used for medicinal use. Despite the plentiful seed production, skilled aren't many seedlings in the wild, that led to a all-encompassing investigation on its generative biology. As a dioecious liana, female plants can produce crop even when there are no nearby male plants, and even the cought, unopened flower buds  eventually mature into seeds. A secret development is connected to the lack of pollinators, airborne irritant grains, and the production of a distinct seed from a tricarpillary gynoecium. In consequence of curiosity, flower and product development, studies on irritant viability, irritant germination, pollen beginning ratio and embryological studies etc were transported. DNA isolation was approved using the leaves of mother plant and seedlings bred from the same plant. High quality DNA was acquired from all the explants using the Modified CTAB plan. The samples were diluted and ISSR reasoning was carried out using 10 ISSR primers. Regardless of the lack of irritant on the stigma and irritant tube seepage into the stylar tube, embryo growth was seen in 80% of the long-term microtome sections calm with fruit primordia that were old from the first to third weeks. Genetic similarity was also told by comparative microscopic analyses of the mother plant and the equivalent F1 seedlings, which financed apomixis.

Author(s) Details:

C. R. Chitra,
Jawaharlal Nehru Tropical Botanic Garden and Research Institute, Palode, Thiruvananthapuram, Kerala – 695562, India.

C. AnilKumar,
Jawaharlal Nehru Tropical Botanic Garden and Research Institute, Palode, Thiruvananthapuram, Kerala – 695562, India.

Please see the link here: https://stm.bookpi.org/ARBS-V2/article/view/11642