NDLI: Single step direct transgenic plant regeneration from adventive embryos of agro-infected sugarcane (Saccharum spp.) spindle leaf roll segments with assured genetic fidelity

Content Provider	Springer Nature Link
Author	Sandhu, Jagdeep Singh Kaur, Manpreet Kaur, Ajinder Kalia, Anu
Copyright Year	2016
Abstract	Single step direct transgenic plant regeneration from agro-infected sugarcane (Saccharum spp.) spindle leaf roll segments with assured genetic fidelity is reported. The pre-cultured (96 h) leaf segments of cultivar CoJ 83 were incubated (10 min) with intermittent shaking in Agrobacterium suspension (OD 1.0–1.2) carrying antifungal gene encoding β-1,3-glucanase under the control of CaMV 35S promoter and NOS terminator, in presence of acetosyringone. Thereafter, the segments were co-cultivated (72 h) and transferred for shoot regeneration. The scanning electron micrographs of sectioned leaf segments undergoing in vitro shoot regeneration revealed occurrence of globular structures in clusters near cut leaf edges within 10 days of incubation. In each cluster, two to ten structures of varying sizes occurred concurrently. The sectioning of globular structures confirmed that these were arising directly from pre-embryogenic determined palisade cells, had unicellular origin and were referred to as adventive embryos. The adventive embryos successively expressed their potential for direct shoot regeneration without callus interphase in 3 weeks and transgenic plant formation within a short period of 8 weeks. The integration of β-1,3-glucanase transgene and its expression in T$_{0}$ plants were determined by reverse transcription-PCR and quantitative real time-PCR, respectively. Simple sequence repeat DNA markers did not detect any polymorphism in the transgenic plants, pointing towards assured genetic fidelity of the plants.
Starting Page	149
Ending Page	162
Page Count	14
File Format	PDF
ISSN	01676857
Journal	Plant Cell, Tissue and Organ Culture (PCTOC)
Volume Number	125
Issue Number	1
e-ISSN	15735044
Language	English
Publisher	Springer Netherlands
Publisher Date	2016-01-02
Publisher Place	Dordrecht
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Adventive embryos Agro-infection Palisade cells qRT-PCR Scanning electron microscopy Simple sequence repeat Unicellular origin Plant Sciences Plant Physiology Plant Genetics & Genomics Plant Pathology
Content Type	Text
Resource Type	Article
Subject	Horticulture

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