Physical and chemical mutagenesis in Stevia rebaudiana: variant generation with higher UGT expression and glycosidic profile but with low photosynthetic capabilities

详细信息    查看全文

• 作者：Shamshad Ahmad Khan ; Laiq ur Rahman ; Rajesh Verma…
• 关键词：Mutagen ; EMS ; Gamma radiation ; Stevia rebaudiana ; Steviol glycosides ; HPLC ; RT ; PCR ; Soil analysis
• 刊名：Acta Physiologiae Plantarum
• 出版年：2016
• 出版时间：January 2016
• 年：2016
• 卷：38
• 期：1
• 全文大小：5,869 KB
• 参考文献：Arnon DI (1949) Copper enzymes in isolated chloroplasts: polyphenoloxidase in Beta vulgaris. Plant Physiol 24:1–15PubMedCentral CrossRef PubMed
  Behera M, Panigrahi J, Mishra RR, Rath SP (2012) Analysis of EMS induced in vitro mutants of Asteracantha longifolia (L.) Nees using RAPD markers. Indian J Biotechnol 11:39–47
  Charbaji T, Nabulsi I (1999) Effect of low doses of gamma irradiation on in vitro growth of grapevine. Plant Cell Tissue Organ Cult 57:129–132CrossRef
  Das K, Dang R, Shivananda TN, Sekeroglu N (2007) Influence of bio-fertilizers on the biomass yield and nutrient content in Stevia rebaudiana (Bertoni) grown in the Indian subtropics. J Med Plant Res 1:5–8
  Edwards A, Johnstone C, Thompson C (1991) A simple and rapid method for the preparation of plant genomic DNA for PCR analysis. Nucleic Acids Res 19:1349PubMedCentral CrossRef PubMed
  Hasbullah NA, Taha RM, Saleh A, Mahmad N (2012) Irradiation effect on in vitro organogenesis, callus growth and plantlet development of Gerbera jamesonii. Horticult Brasil 30:252–257CrossRef
  Jain SM, Maluszynski M (2004) Induced mutations and biotechnology on improving crops. In: Mujib A, Cho M, Predieri A, Banerjee S (eds) In vitro applications in crop improvement: recent progress. IBH, Oxford, pp 169–202
  Jain SM, Spencer MM (2006) Biotechnology and mutagenesis in improving ornamental plants in Floriculture and Ornamental Biotechnology. Advances and Tropical Issues. Teixeira da Silva JA (Ed.). Global Science Books Ltd, London 1:1749–2036
  Jain P, Kachhwaha S, Kothari SL (2009) Improved micropropagation protocol and enhancement in biomass and chlorophyll content in Stevia rebaudiana (Bert) Bertoni by using high copper levels in the culture medium. Sci Hort 119:315–319CrossRef
  Jain P, Kachhwaha S, Kothari SL (2014) Chloroplast ultrastructure, photosynthesis and enzyme activities in regenerated plants of Stevia rebaudiana (Bert.) Bertoni. Indian J Exp Biol 52:898–904PubMed
  Kalia RK, Rai MK, Kalia S, Singh R, Dhawan AK (2011) Microsatellite markers: an overview of the recent progress in plants. Euphytica 177:309–334CrossRef
  Khan SA, Rahman L, Shanker K, Singh M (2014) Agrobacterium tumefaciens-mediated transgenic plant and somaclone production through direct and indirect regeneration from leaves in Stevia rebaudiana with their glycoside profile. Protoplasma 251:661–670CrossRef PubMed
  Kim JH, Baek MH, Chung BY, Wi SG, Kim JS (2004) Alterations in the photosynthetic pigments and antioxidant machineries of red pepper (Capsicum annum L.) seedlings from gamma-irradiated seeds. J Plant Biol 47:314–321CrossRef
  Kovacs E, Keresztes A (2002) Effect of gamma and UV-B/C radiation on plant cell. Micron 33:199–210CrossRef PubMed
  Kumar H, Kaul K, Gupta SB, Kaul VK, Kumar S (2012) A comprehensive analysis of fifteen genes of steviol glycosides biosynthesis pathway in Stevia rebaudiana (Bertoni). Gene 492:276–284CrossRef PubMed
  Kura-Hotta M, Satoh K, Katoh S (1987) Relationship between photosynthesis and chlorophyll content during leaf senescence of rice seedlings. Plant Cell Physiol 28:1321–1329
  Luan YS, Zhang J, Gao XR, An LJ (2007) Mutation induced by ethylmethylsulfonate (EMS), in vitro screening for salt tolerance and plant regeneration of sweet potato (Ipomoea batatas L.). Plant Cell Tissue Organ Cult 88:77–81CrossRef
  McCallum CM, Comai L, Geene EA, Henikoff S (2000) Targeted screening for induced mutations. Nat Biotechnol 18:455–457CrossRef PubMed
  Megeji NW, Kumar JK, Singh V, Kaul VK, Ahuja PS (2005) Introducing Stevia rebaudiana. A natural Zero-Calorie sweetener. Current Sci 88:801–804
  Monteiro WR, Castro MM, Mazzoni-Viveiros CS, Mahlberg PG (2001) Development and some histochemical aspects of foliar glandular trichomes of Stevia rebaudiana (Bert.) Bert. Asteraceae. Revta Brasil Bot 24:349–357
  Nakai T, Konishi T, Zhang XQ, Chollet R, Tonouchi N, Tsuchida T, Yoshinga F, Mori H, Sakai F, Hayashi T (1998) An increase in apparent affinity for sucrose of mung bean sucrose synthase is caused by in vitro phosphorylation or directed mutagenesis of Ser. Plant Cell Physiol 39:1337–1341CrossRef PubMed
  Nei M, Li WH (1979) Mathematical model for studying genetic variation in terms of restriction endonucleases. PNAS 76:5269–5273PubMedCentral CrossRef PubMed
  Nelson DW, Sommers LE(1996) Total carbon, organic carbon, and organic matter. In Sparks DL (ed.) Methods of Soil Analysis, Part 3, Chemical Methods. SSSA Book Series No. 5. Soil Science Society of American Madison, WI. pp 153–188
  Oliveira VM, Forni-Martins ER, Magalhaes PM, Alves MN (2004) Chromosomal and morphological studies of diploid and polyploid cytotypes of Stevia rebaudiana (Bertoni) (Eupatorieae, Asteraceae). Gen Mol Biol 27:215–222CrossRef
  Richman A, Swanson A, Humphrey T, Chapman R, McGarvey B, Pocs R, Brandle J (2005) Functional genomics uncovers three glucosyltransferases involved in the synthesis of the major sweet glucosides of Stevia rebaudiana. Plant J 41:56–67CrossRef PubMed
  Sivaram L, Mukundan U (2003) In vitro culture on Stevia rebaudiana. In Vitro Cell Develop Biol Plant 39:520–523CrossRef
  Subbiah BV, Asija GL (1956) A rapid procedure for estimation of available nitrogen in soils. Curr Sci 25:259–260
  Venkataiah P, Christopher T, Karampuri S (2005) Selection of atrazine resistant plants by in vitro mutagenesis in pepper (Capsicum annuum). Plant Cell Tiss Organ Cult 83:75–82CrossRef
  Verma P, Mathur AK, Shankar K (2012) Enhanced vincamine production in selected tryptophan-overproducing shoots of Vinca minor. Plant Cell Tiss Organ Cult 111:239–245CrossRef
  Wi SG, Chung BY, Kim JH, Baek MH, Yang DH, Lee JW, Kim JS (2005) Ultrastructural changes of cell organelles in Arabidopsis stem after gamma irradiation. J Plant Biol 48:195–200CrossRef
  Yadav SK, Guleria P (2012) Steviol glycosides from Stevia: biosynthesis pathway review and their application in foods and medicine. Crit Rev Food Sci Nutri 52:988–998CrossRef
  
• 作者单位：Shamshad Ahmad Khan (1)
  Laiq ur Rahman (1)
  Rajesh Verma (2)
  Karuna Shanker (3)
  
  1. Plant Biotechnology Division, Central Institute of Medicinal and Aromatic Plants (CSIR-CIMAP), Kukrail Picnic Spot Road, P.O. CIMAP, Lucknow, 226015, India
  2. Department of Soil Analysis, Central Institute of Medicinal and Aromatic Plants (CSIR-CIMAP), Kukrail Picnic Spot Road, P.O. CIMAP, Lucknow, 226015, India
  3. Analytical Chemistry Division, Central Institute of Medicinal and Aromatic Plants (CSIR-CIMAP), Kukrail Picnic Spot Road, P.O. CIMAP, Lucknow, 226015, India
  
• 刊物主题：Plant Physiology; Plant Genetics & Genomics; Plant Biochemistry; Plant Pathology; Plant Anatomy/Development; Agriculture;
• 出版者：Springer Berlin Heidelberg
• ISSN：1861-1664

文摘

The leaf explants of Stevia rebaudiana were exposed to a range of EMS (chemical mutagen) and gamma radiation (physical mutagen) doses. Among all the treatments, doses of 0.4 % v/v EMS and 0.95 KR gamma radiation were found to be most effective treatment for selection of variants via direct shoot bud induction. The field-established plants designated as EMS (E), gamma (G), and control (C) plants were subjected to photosynthetic parameter studies where E and G plants showed lower chlorophyll content, transpiration rate, CO₂ exchange rate, and stomatal conductance. The implanted field soil at the time of harvesting of plants showed reduction in organic carbon (OC), electrical conductivity (EC), phosphorus (P)/potassium (K), and total Kjeldahl nitrogen (TKN). The ISSR profiling of E, G, and C scored total of 107 bands out of which 63 % were polymorphic bands, and plant E was found to be more distant from the C plant phylogenetically. On phytochemical analysis, the G plant registered twofold enhanced rebaudioside A with lower stevioside (3.2 ± 0.22 % dry wt.) content, whereas E plant showed more than 1.5- and 2.0-fold increase in stevioside and rebaudioside A as compared to control plants. Both the E (3.1 ± 0.15 % dry wt.) and G (2.3 ± 0.21 % dry wt.) plants registered lower steviol. The enhanced steviol glycoside profile was supported by the RT-PCR analysis of UGT74G1 and UGT76G1 that corresponds to stevioside and rebaudioside A biosynthesis, respectively. While the E plant showed 5–6-fold increase in the UGT74G1 (RQ = 5.51 ± 0.5) and UGT76G1 (RQ = 6.61 ± 0.5) gene expression, the G plant showed 5-fold increase in UGT76G1 (RQ = 5.29 ± 0.2) gene expression. Keywords Mutagen EMS Gamma radiation Stevia rebaudiana Steviol glycosides HPLC RT-PCR Soil analysis