Skip to page content
USDA Forest Service
  
Treesearch

Research & Development Treesearch

 
Treesearch Home
About Treesearch
Contact Us
Research & Development
Forest Products Lab
International Institute of Tropical Forestry
Northern
Pacific Northwest
Pacific Southwest
Rocky Mountain
Southern Research Station
Help
 

Science.gov - We Participate


USA.gov  Government Made Easy


Global Forest Information Service

US Forest Service
P.O. Box 96090
Washington, D.C.
20090-6090

(202) 205-8333

You are here: Home / Search / Publication Information
Bookmark and Share

Publication Information

View PDF (1.0 MB byte)

Title: Protoplast isolation and genetically true-to-type plant regeneration from leaf- and callus-derived protoplasts of Albizia julibrissin

Author: Rahmani, Mohammad-Shafie; Pijut, Paula M.; Shabanian, Naghi;

Date: 2016

Source: Plant Cell, Tissue and Organ Culture (PCTOC). 127(2): 475-488.

Publication Series: Scientific Journal (JRNL)

Description: Protoplast isolation and subsequent plant regeneration of Albizia julibrissin was achieved from leaf and callus explants. Leaf tissue from 4 to 5-week-old in vitro seedlings was the best source for high-yield protoplast isolation. This approach produced 7.77 × 105 protoplasts (Pp) per gram fresh weight with 94 % viability; after 60 min pre-plasmolysis with 0.7 M sorbitol followed by digestion in a solution of cell and protoplast wash plus 0.7 M mannitol, 1.5 % cellulase Onozuka R10, and 1 % pectolyase Y-23 for 6 h. Liquid Kao and Michayluk medium containing 2.7 μM α-naphthaleneacetic acid (NAA) and 2.2 μM 6-benzylaminopurine (BA) was best for sustained cell division and microcolony formation from both leaf- and callus-derived protoplasts at a density of 3–5 × 105 Pp ml−1. Protoplast-derived microcalli became visible after 3–4 weeks on semi-solid medium of the same composition. Microcalli were then cultured on Murashige and Skoog (MS) medium containing Gamborg B5 vitamins or woody plant medium supplemented with different concentrations of NAA plus 4.4 μM BA for further growth. Proliferated leaf- and callus-protoplast-derived calli differentiated into microshoots on MS medium containing 13.2 μM BA plus 4.6 μM zeatin after 2–3 weeks, with an overall shoot organogenesis efficiency of 78–93 %. Rooting of microshoots on half-strength MS medium containing 4.9 μM indole-3-butyric acid was successful, and plantlets were acclimatized to the greenhouse with a survival rate of >62 %. Using ten start codon targeted and ten inter-simple sequence repeat primers, the genetic integrity of nine leafand six callus-protoplast-based plants was validated along with the mother seedlings.

Keywords: Albizia, Cellulase, CPW, Genetic fidelity, Microcolony formation, Protoplast, Silk tree

Publication Notes:

  • We recommend that you also print this page and attach it to the printout of the article, to retain the full citation information.
  • This article was written and prepared by U.S. Government employees on official time, and is therefore in the public domain.
  • This publication may be available in hard copy. Check the Northern Research Station web site to request a printed copy of this publication.
  • Our on-line publications are scanned and captured using Adobe Acrobat. During the capture process some typographical errors may occur. Please contact Sharon Hobrla, shobrla@fs.fed.us if you notice any errors which make this publication unusable.

XML: View XML

Citation:


Rahmani, Mohammad-Shafie; Pijut, Paula M.; Shabanian, Naghi. 2016. Protoplast isolation and genetically true-to-type plant regeneration from leaf- and callus-derived protoplasts of Albizia julibrissin. Plant Cell, Tissue and Organ Culture (PCTOC). 127(2): 475-488.

 


 [ Get Acrobat ]  Get the latest version of the Adobe Acrobat reader or Acrobat Reader for Windows with Search and Accessibility

USDA logo which links to the department's national site. Forest Service logo which links to the agency's national site.