Seed-derived Plants Research Articles

Determination for inflorescence development is a stable state, separable from determination for flower development in Pisum sativum L. buds

Terminal meristems of Pisum sativum (garden pea) transit from vegetative to inflorescence development, and begin producing floral axillary meristems. Determination for inflorescence development was assessed by culturing excised buds and meristems. The first node of floral initiation (NFI) for bud expiants developing in culture and for adventitious shoots forming on cultured meristems was compared with the NFI of intact control buds. When terminal buds having eight leaf primordia were excised from plants of different ages (i.e., number of unfolded leaves) and cultured on 6-benzylaminopurine and kinetin-supplemented medium, the NFI was a function of the age of the source plant. By age 3, all terminal buds were determined for inflorescence development. Determination occurred at least eight nodes before the first axillary flower was initiated. Thus, the axillary meristems contributing to the inflorescence had not formed at the time the bud was explanted. Similar results were obtained for cultured axillary buds. In addition, meristems excised without leaf primordia from axillary buds three nodes above the cotyledons of age-3 plants gave rise to adventitious buds with an NFI of 8.3 ±0.3 nodes. In contrast seed-derived plants had an NFI of 16.5 ±0.2. Thus cells within the meristem were determined for inflorescence development. These findings indicate that determination for inflorescence development in P. sativum is a stable developmental state, separable from determination for flower development, and occurring prior to initiation of the inflorescence at the level of meristems.

Indirect somatic embryogenesis and plant recovery from cotton (Gossypium hirsutum L.)

We describe a tissue culture procedure for somatic embryogenesis and plantlet regeneration in cotton (Gossypium hirsutum L. cv. Coker 312). Callused explants or individual globular embryos were transferred to basal media to induce somatic embryogenesis. To determine characteristic early indicators of successful germination and conversion, we identified six types of embryos that developed on basal media. Two of the six embryo types, designated as tulip-shaped and trumpet-shaped, could undergo conversion in preliminary tests, whereas the others had little or no developmental potential. Several media treatments designed to enhance the maturation of globular somatic embryos failed to increase the fraction of embryos which matured to form recoverable types. In efforts to improve plantlet recovery, tulip-shaped embryos were used in limited trials to contrast the effects of chemical and physical desiccation treatments on germination and conversion. The selective use of tulip-shaped somatic embryos, coupled with partial desiccation, seems to have augmented plant recovery. Growth habit, flowering, seed set, and lint production of most of the regenerated plants were comparable to seed-derived plants grown under the same conditions.

Efficient plant regeneration from rice protoplasts in general medium

We established an efficient and reproducible procedure for protoplast propagation and fertile plant regeneration of rice (Oryza sativa L.) cultivars Nipponbare and Taipei 309. Selection of scutellum-derived secondary calli, the use of General medium and nurse culture were all found to be critical in the procedure. When 5 basal media (Murashige and Skoog, RY-2, modified R2, Amino Acid and General media) were compared, suspension callus growth rate, protoplast yield and plating efficiency were all about 30% higher in General medium than in the second-best R2 medium. Only one month was required to develop suspension cultures for protoplast isolation using General medium. A plating efficiency as high as 17% and a plant regeneration frequency of 67% were achieved by the improved procedure. Agronomic traits of protoplast- and seed-derived plants were found to be similar.

Plant regeneration from somatic embryogenic suspension cultures of cotton (Gossypium hirsutum L.)

Maintainable, highly embryogenic suspension cultures of cotton (Gossypium hirsutum L. cv. 'Coker 310') have been obtained. Callus cultures were initiated from cotyledonary tissues from aseptically-germinated seedlings. To establish the suspension cultures, callus tissue was placed in a liquid medium containing either 0.5 mg/l picloram or 0.1 mg/l 2,4-dichlorophenoxyacetic acid. For proliferation of the embryogenic suspension, 5 mg/l of 2,4-dichlorophenoxyacetic acid was used. Embryo development took place when the embryogenic tissue was transferred to an auxin-free liquid medium containing 15 mM glutamine. Early embryo development was fairly synchronous and large numbers of somatic embryos were produced. Regenerated plants were fertile and smaller than seed-derived plants.

Concluding Remarks

Abstract Potential applications of synthetic seed technology to agriculture are numerous, but certain milestones are ahead. The driving force behind commercialization must be the added benefit conferred by use of synthetic seed-derived plants, and this benefit will vary greatly in magnitude depending on specific applications. Increased crop value must be great enough to justify new research and development costs that are not encountered in traditional seed production. Anticipated profit for a few high-value applications will fuel basic advances in technology that will be adaptable to many other crops and applications.

Floral determination in axillary buds of Nicotiana silvestris

At anthesis of the terminal flower the developmental fates of axillary buds of the long-day plant Nicotiana silvestris were assessed in situ and in isolation. The in situ developmental fate was assessed by decapitating the plant above the bud in question and letting the bud mature. The developmental fate of isolated buds was assessed by removing the bud from the main axis, rooting it, and letting it mature. The number of nodes below the terminal flower of the mature shoot was indicative of the developmental fate of the bud. Terminal meristems of rooted axillary buds exhibited two patterns of development: (1) Their developmental fate was the same as that of in situ buds at the same node or (2) their developmental fate was the same as that of seed-derived plants. For example, terminal meristems of rooted buds from the fourth node below the inflorescence produced either 15 to 19 nodes or 36 to 40 nodes. In situ fourth buds produced 12 to 14 nodes while seed-derived plants produced 33 to 39 nodes. Terminal meristems of rooted axillary buds that exhibited the same developmental fate as that of in situ buds were determined for floral development. Although determined buds produced a terminal flower, all but one had abnormal inflorescences. That is, in the place of floral branches determined buds produced vegetative branches. Four buds that were not determined for floral development had their shoot tips rooted each time the plant bolted. Only when the plants were allowed to grow without being rerooted did they flower. These results indicate that roots may prevent and/or destabilize floral determination in N. silvestris.

The use of genetic markers selected in vitro for the isolation and genetic verification of intraspecific somatic hybrids of Nicotiana tabacum L.

A scheme employing genetic markers obtained by in vitro selection was developed for the stringent isolation of hybrid somatic cells of Nicotiana tabacum. Mesophyll protoplasts that carried two dominant alleles of nuclear genes conferring resistance to the herbicide picloram (pmR1) and the ability to utilize glycerol as the sole source of carbon (Gut) were fused with suspension-culture protoplasts that were marked with the dominant nuclear allele (HuR9) conferring resistance to hydroxyurea. Putative somatic hybrid cell lines were identified by selecting for the Gut and HuR9 markers, followed by an assay for the unselected marker PmR1. Plants regenerated from six of these cell lines were proved to be true somatic hybrids by demonstrating the segregation of each of the three parental markers in the progeny of crosses of those plants with normal seed-derived plants.

Greenhouse propagation of Cannabis Sativa L. by vegetative cuttings

Previous work revealed significant variations in cannabinoid profiles ofCannabis sativa L. derived from a single seed source (P.1. 378939) and subjected to the same growth environment. Studies were conducted to evaluate the efficacy of propagation ofC. sativa by vegetative cuttings in order to increase uniformity of cannabinoid concentrations within a given plant population.C. sativa was successfully propagated by vegetative cuttings. However, there were both morphological and biochemical differences between seed-derived plants and their vegetative propagules. Delta-9-tetrahydrocannabinol concentrations were 4.1 times higher in vegetative propagules than in seed propagules. Vegetative cuttings also generally developed more profuse lateral branch growth; hence, foliage increased relative to their parent plants. Cannabinoid levels within the population of vegetative cuttings remained highly variable.