Indirect somatic embryogenesis and morphohistological analysis in Capsicum chinense

Abstract

Capsicum chinense is recalcitrant in in vitro morphogenesis. No efficient, reproducible somatic embryogenesis regeneration system exists for this species, impeding regeneration from transformed cells. An indirect somatic embryogenesis protocol is developed using mature C. chinense zygotic embryo segments (ZES). The ZES cultured in semi-solid Murashige-Skoog (MS) medium supplemented with 8.9 µM naphthaleneacetic acid, 11.4 µM indoleacetic acid and 8.9 µM 6-benzylaminopurine, developed an embryogenic callus and 8% of the calli developed somatic embryos. Torpedo-stage somatic embryos were detached from the callus and subcultured in semi-solid MS medium without growth regulators, producing a 75% conversion rate to plantlets with well-formed root tissue. Histological analysis showed the developed structures to have no vascular connection with the callus and to be bipolar, confirming that this protocol induced formation of viable somatic embryos from mature C. chinense ZES. All acclimated plantlets survived under greenhouse conditions. This protocol will facilitate regeneration of genetically transformed plants using either biolistics or Agrobacterium tumefaciens approach.

Somatic embryogenesis and plant regeneration of Capsicum baccatum L.

“…Auxins and cytokinin are the main plant growth regulators involved in the regulation of plant cell cycling, division and differentiation, playing key roles in somatic embryogenesis of many species [21], [22], [23], [24] including Capsicum species [13], [14], [18], [25]. In the present study, asynchronous development of somatic embryos was noticed as reported earlier in C. annuum [25], [26], [27], [28] and C. chinense [29].…”

Somatic Embryogenesis: Fundamental Aspects and Applications

“…Ace as explants, Buyukalaca and Mavituna (1996) reported indirect somatic embryogenesis on MS medium supplemented with 9.05 μM 2,4-D and 3 % sucrose, where embryogenic callus was generated and then subcultured in MS liquid medium containing 4.52 μM 2,4-D and 3 % sucrose to increase the embryogenic callus mass as a suspension culture. Solís-Ramos et al (2010) established a protocol for the induction of indirect somatic embryogenesis in C. chinense starting with mature zygotic embryos segments as explants, which were cultured on MS medium supplemented with 8.9 μM NAA, 11.4 μM IAA, and 8.6 μM BA to produce embryogenic callus; only 8 % of calli produced somatic embryos and when torpedo-stage somatic embryos were detached from callus and cultured on MS medium without growth regulators a 75 % conversion into plantlets was attained; these plants were finally transplanted to soil and adapted to greenhouse conditions. Maturation of somatic embryos and conversion into plants was achieved at 97 % efficiency on paper bridges in a half-strength MS medium with 1.89 μM ABA.…”

Plant regeneration, developmental pattern and genetic fidelity of somatic embryogenesis derived Musa spp.

“…The study showed that most of the suspensions were single cells with some multi-celled embryogenic clump indicating asynchronous induction at different stages of somatic embryos. Non-embryogenic cells are characterized by large vacuole in the callus cells indicating cell degradation, as vacuole plays a critical role in programmed cell death [36]. The acquisition of embryogenic competence has been attributed to the cells that show meristematic traits during the induction phase [37].The present histological study demonstrates the presence of all important stages during somatic embryogenesis in banana and importance of utilizing the optimal stage for promoting embryogenic competence.…”