True-to-type micropropagated plants of para rubber (Hevea brasiliensis Müll. Arg.) via somatic embryogenesis
Keywords:DNA fingerprinting; para rubber; RAPD; SCoT; SSRs
Plant micropropagation via somatic embryogenesis is a powerful technique for rapid mass propagation, especially in para rubber (Hevea brasiliensis Müll. Arg.). However, somaclonal variations are the major limitation of this process. To date, DNA fingerprinting, i.e., RAPD (Randomly Amplified Polymorphic DNA), Star Codon Targeted (SCoT), and SSRs (Simple Sequence Repeats), is one of the most successful technologies to detect the genetic fidelity in the somatic embryos. The aim of present study was to induce somatic embryos from inner integument explants of para rubber cv. ‘RRIM 600’ at different developmental stages and subsequent acclimatization and transplantation (under greenhouse and field conditions) of the propagated seedlings. The genetic stability of the plants derived from somatic embryos was also analysed in comparison to the mother plant using RAPD, SCoT and SSRs markers. Somatic embryos derived from inner integuments of 5-week-old immature seeds after pollination were more efficient than older and younger seeds. In addition, para rubber mother plants cv. ‘RRIM600’ and plants derived from somatic embryogenesis demonstrated the same pattern of DNA fragments, as confirmed by three PCR-based techniques, RAPD, SCoT and SSRs, whereas these in the pattern were different from ‘RRIT 226’, ‘PB 235’, ‘PB 251’, ‘PB 255’ and ‘BMP 24’. Interestingly, T2 plant was found to possess somaclonal variations when compared with mother plant. Based on the results, we confirm that the plants derived from somatic embryogenesis of para rubber cv. ‘RRIM 600’ were true-to-type to that of ‘RRIM 600’ master stock.
Agarwal T, Gupta AK, Patel AK, Shekhawat NS (2015). Micropropagation and validation of genetic homogeneity of Alhagi maurorum using SCoT, ISSR and RAPD markers. Plant Cell, Tissue and Organ Culture 120:313-323. https://doi.org/10.1007/s11240-014-0608-z
Anandan R, Prakash M, Deenadhayalan T, Nivetha R, Kumar NS (2018). Efficient in vitro plant regeneration from cotyledon-derived callus cultures of sesame (Sesamum indicum L.) and genetic analysis of true-to-type regenerants using RAPD and SSR markers. South African Journal of Botany 119:244-251. https://doi.org/10.1016/j.sajb.2018.09.020
Bhattacharyya P, Kumaria S, Bose B, Paul P, Tandon P (2017a). Evaluation of genetic stability and analysis of phytomedicinal potential in micropropagated plants of Rumex nepalensis–A medicinally important source of pharmaceutical biomolecules. Journal of Applied Research on Medicinal and Aromatic Plants 6:80-91. https://doi.org/10.1016/j.jarmap.2017.02.003
Bhattacharyya P, Kumar V, van Staden J (2017b). Assessment of genetic stability amongst micropropagated Ansellia africana, a vulnerable medicinal orchid species of Africa using SCoT markers. South African Journal of Botany 108:294-302. https://doi.org/10.1016/j.sajb.2016.11.007
Blanc G, Lardet L, Martin A, Jacob JL, Carron MP (2002). Differential carbohydrate metabolism conducts morphogenesis in embryogenic callus of Hevea brasiliensis (Müll. Arg.). Journal of Experimental Botany 53:1453-1462. https://doi.org/10.1093/jexbot/53.373.1453
Bose B, Kumaria S, Choudhury H, Tandon P (2016). Assessment of genetic homogeneity and analysis of phytomedicinal potential in micropropagated plants of Nardostachys jatamansi, a critically endangered, medicinal plant of alpine Himalayas. Plant Cell, Tissue and Organ Culture 124:331-349. https://doi.org/10.1007/s11240-015-0897-x
Bradaï F, Sánchez-Romero C, Martín C (2019). Somaclonal variation in olive (Olea europaea L.) plants regenerated via somatic embryogenesis: Influence of genotype and culture age on genetic stability. Scientia Horticulturae 251:260-266. https://doi.org/10.1016.j.scienta.2019.03.010
Bychappa M, Mishra MK, Jingade P, Huded AK (2019). Genomic alterations in coding region of tissue culture plants of Coffea arabica obtained through somatic embryogenesis revealed by molecular markers. Plant Cell, Tissue and Organ Culture 139:91-103. https://doi.org/10.1007/s11240-019-01666-8
Cardinal ÁBB., Gonçalves PDS., Martins, ALM (2007). Stock-scion interactions on growth and rubber yield of Hevea brasiliensis. Scientia Agricola 64:235-240. http://dx.doi.org/10.1590/S0103-90162007000300004
Carron MP, Enjalric F, Lardet L, Deschamps A (1989). Rubber (Hevea brasiliensis Müll. Arg.). In: Bajaj YPS (Eds.), Biotechnology in Agriculture and Forestry, vol. 5 Trees II, Springer, Berlin, Heidelberg. pp 222-245.
Chantuma P, Lacote R, Leconte A, Gohet E (2011). An innovative tapping system, the double cut alternative, to improve the yield of Hevea brasiliensis in Thai rubber plantations. Field Crops Research 121:416-422. https://doi.org/10.1016/j.fcr.2011.01.013
Cornish K (2017). Alternative natural rubber crops: why should we care? Technology and Innovation 18:244-255. https://doi.org/10.21300/18.4.2017.245
Golbon R, Cotter M, Sauerborn J (2018). Climate change impact assessment on the potential rubber cultivating area in the Greater Mekong Subregion. Environmental Research Letters 13:084002.
Gonçalves PS, Martins ALM (2002). Combining ability effects of clonal rootstocks and scions in rubber trees (Hevea). Crop Breeding and Applied Biotechnology 2:445-452.
Gonçalves PS, Scaloppi Jr EJ, Martins MA, Moreno RMB, Branco RBF, Gonçalves ECP (2011). Assessment of growth and yield performance of rubber tree clones of the IAC 500 series. Pesquisa Agropecuária Brasileira 46:1643-1649. https://doi.org/10.1590/S0100-204X2011001200009
Haque SM, Ghosh B (2016). High-frequency somatic embryogenesis and artificial seeds for mass production of true-to-type plants in Ledebouria revoluta: An important cardioprotective plant. Plant Cell, Tissue and Organ Culture 127:71-83. https://doi.org/10.1007/s11240-016-1030-5
Hazir MHM, Kadir RA, Gloor E, Galbraith D (2020). Effect of agroclimatic variability on land suitability for cultivating rubber (Hevea brasiliensis) and growth performance assessment in the tropical rainforest climate of Peninsular Malaysia. Climate Risk Management 27:100203. https://doi.org/10.1016/j.crm.2019.100203
IRSG (International Rubber Study Group) (2020). Statistical Summary of World Rubber Situation (Singapore: International Rubber Study Group) (https://www.statista.com/statistics/275390/world-usage-distribution-of-natural-rubber/)
Jaccard P (1901). Étude comparative de la distribution florale dans une portion des Alpes et des Jura. Bull Soc Vandoise Sci Nat 37:547-579.
Kalawong S, Srichuay W, Sirisom Y, Te-chato S (2014). Microcutting as a tool for propagation and genetic transformation in rubber tree. Thaksin University Journal 17:15-25.
Karumamkandathil R, Uthup TK, Sankaran S, Unnikrishnan D, Saha T, Nair SS (2015). Genetic and epigenetic uniformity of polyembryony derived multiple seedlings of Hevea brasiliensis. Protoplasma 252:783-796. https://doi.org/10.1007/s00709-014-0713-1
Kouassi KM, Koffi KE, Gnagne M, Kone M, Kouakou TH (2013). Influence of plant growth regulators on somatic embryogenesis induction from inner teguments of rubber (Hevea brasiliensis) seeds. African Journal of Biotechnology 12:1972-1977. https://doi.org/10.5897/AJB12.2411
Lardet L, Dessailly F, Carron MP, Montoro P, Monteuuis O (2008). Influences of aging and cloning methods on the capacity for somatic embryogenesis of a mature Hevea brasiliensis genotype. Tree Physiology 29:291-298. https://doi.org/10.1093/treephys/tpn027
Lema-Rumińska J, Kulus D, Tymoszuk A, Varejão JM, Bahcevandziev K (2019). Profile of secondary metabolites and genetic stability analysis in new lines of Echinacea purpurea (L.) Moench micropropagated via somatic embryogenesis. Industrial Crops and Products 142:111851. https://doi.org/10.1016/j.indcrop.2019.111851
Liyanage KK, Sumanasinghe VA, Attanayake DPSTG, Baddewithana BWAN (2014). Identification of recommended Hevea brasiliensis (Willd. ex A. Juss.) Müll. Arg. clones grow in Sri Lanka by RAPD analysis. Tropical Agricultural Research 25:188-200.
Masson A, Julien JM, Boedt L (2013). Industrial propagation by rooted cuttings of mature selected clones of Hevea brasiliensis. Bois et Forêts des Tropiques 317:51-58.
Masson A, Monteuuis O (2016). Mass production of self-rooted Hevea brasiliensis industrial clones by tissue culture and nursery methods. Proceeding of the Fourth International Conference of the IUFRO Unit 2.09.02 on Development and application vegetative propagation technologies in plantation forestry to cope with a changing climate and environment. September 19-23, 2016, La Plata, Argentina.
Masson A, Monteuuis O (2017). Rubber tree clonal plantations: grafted vs self-rooted plant material. Bois et Forêts des Tropiques 332:58-68.
Modeste KK, Edmond KK, Konkon NÃ, Miche, G, Hilaire KT (2012). Callogenesis and somatic embryogenesis induction in Hevea brasiliensis: Effects of fruit shelf-life and carbon source. Research in Biotechnology 3:42-50.
Muthukumar M, Kumar TS, Rao MV (2016). Organogenesis and evaluation of genetic homogeneity through SCoT and ISSR markers in Helicteres isora L., a medicinally important tree. South African Journal of Botany 106:204-210. https://doi.org/10.1016/j.sajb.2016.07.017
Nakkanong K, Nualsri C, Sdoodee S (2008). Analysis of genetic diversity in early introduced clones of rubber tree (Hevea brasiliensis) using RAPD and microsatellite markers. Songklanakarin Journal of Science and Technology 30:553-560.
Nasri F, Zakizadeh H, Vafaee Y, Mozafari AA (2018). Callus induction and plant regeneration of Chrysanthemum morifolium and C. coccineum via direct and indirect organogenesis and genetic fidelity analysis using IRAP, ISSR and SCoT molecular markers. Journal of Ornamental Plants 8:265-284.
Prado MJ, Rodriguez E, Rey L, González MV, Santos C, Rey M (2010). Detection of somaclonal variants in somatic embryogenesis-regenerated plants of Vitis vinifera by flow cytometry and microsatellite markers. Plant Cell, Tissue and Organ Culture 103:49-59. https://doi.org/10.1007/s11240-010-9753-1
Pethin D, Nakkanong K, Nualsri C (2015). Performance and genetic assessment of rubber tree clones in Southern Thailand. Scientia Agricola 72:306-313. https://doi.org/10.1590/0103-9016-2014-0354
Priyadarshan PM, Hoa TT, Huasun H, Gonçalves PS (2005). Yielding potential of rubber (Hevea brasiliensis) in suboptimal environments. Journal of Crop Improvement 14:221-247. https://doi.org/10.1300/J411v14n01_10
Rai MK, Phulwaria M, Gupta AK, Shekhawat NS, Jaiswal U (2012). Genetic homogeneity of guava plants derived from somatic embryogenesis using SSR and ISSR markers. Plant Cell, Tissue and Organ Culture 111:259-264. https://doi.org/10.1007/s11240-012-0190-1
Rahman MM, Mahmood M, Abdullah N, Shaharuddin NA, Parvin W (2017). Somatic embryogenesis and subsequent plant regeneration from zygotic embryo derived callus of rubber (Hevea brasiliensis Müell. Arg). Plant Tissue Culture and Biotechnology 27:51-61.
Rathore MS, Mastan SG, Yadav P, Bhatt VD, Shekhawat NS, Chikara J (2016). Shoot regeneration from leaf explants of Withania coagulans (Stocks) Dunal and genetic stability evaluation of regenerates with RAPD and ISSR markers. South African Journal of Botany 102:12-17. https://doi.org/10.1016/j.sajb.2015.08.003
Ratnasingam J, Ramasamy G, Wai LT, Senin AL, Muttiah N (2015). The prospects of rubberwood biomass energy production in Malaysia. BioResources 10:2526-2548.
Rohela GK, Jogam P, Bylla P, Reuben C (2019). Indirect regeneration and assessment of genetic fidelity of acclimated plantlets by SCoT, ISSR, and RAPD markers in Rauwolfia tetraphylla L.: An endangered medicinal plant. BioMed Research International 2019:3698742. https://doi.org/10.1155/2019/3698742
Rohela GK, Jogam P, Mir MY, Shabnam AA, Shukla P, Abbagani S, Kamili AN (2020). Indirect regeneration and genetic fidelity analysis of acclimated plantlets through SCoT and ISSR markers in Morus alba L. cv. Chinese white. Biotechnology Reports 25:e00417. https://doi.org/10.1016/j.btre.2020.e00417
Sathish D, Vasudevan V, Theboral J, Elayaraja D, Appunu C, Siva R, Manickavasagam M (2018). Efficient direct plant regeneration from immature leaf roll explants of sugarcane (Saccharum officinarum L.) using polyamines and assessment of genetic fidelity by SCoT markers. In Vitro Cellular and Developmental Biology-Plant 54:399-412. https://doi.org/10.1007/s11627-018-9910-5
Savita, Pati PK, Virk GS, Nagpal A (2015). An efficient somatic embryogenesis protocol for Citrus jambhiri and assessment of clonal fidelity of plantlets using RAPD markers. Journal of Plant Growth Regulation 34:309-319. https://doi.org/10.1007/s00344-014-9465-6
Sharma U, Rai MK, Shekhawat NS, Kataria V (2019). Genetic homogeneity revealed in micropropagated Bauhinia racemosa Lam. using gene targeted markers CBDP and SCoT. Physiology and Molecular Biology of Plants 25:581-588. https://doi.org/10.1007/s12298-018-00639-z
Sirisom Y, Te-chato S (2014). Assessment of somaclonal variations of in vitro-plants derived from nodal culture of rubber trees by SSR markers. Songklanakarian Journal of Plant Science 1:7-12.
Srichuay W, Kalawong S, Sirisom Y, Te-chato S (2014). Callus induction and somatic embryogenesis from anther cultures of Hevea brasiliensis Müell Arg. Kasetsart Journal (Natural Science) 48:364-375.
Srichuay W, Te-chato S (2015). Analyzing somaclonal variation in somatic embryo derived from in vitro anther culture of rubber tree (Hevea brasiliensis Müell. Arg.) by simple sequence repeat (SSR) marker and flow cytometry. Songklanakarin Journal of Plant Science 2:27-31.
Supriya R, Priyadarshan PM (2019). Genomic technologies for Hevea breeding. Advances in Genetics 104:1-73. https://doi.org/10.1016/bs.adgen.2019.04.001
Sushamakumari S, Asokan MP, Anthony P, Lowe KC, Power JB, Davey MR (2000). Plant regeneration from embryogenic cell suspension-derived protoplasts of rubber. Plant Cell, Tissue and Organ Culture 61:81-85. https://doi.org/10.1023/A:1006494404224
Thakur J, Dwivedi MD, Sourabh P, Uniyal PL, Pandey AK (2016). Genetic homogeneity revealed using SCoT, ISSR and RAPD markers in micropropagated Pittosporum eriocarpum Royle-An endemic and endangered medicinal plant.PLoS ONE 11:e0159050. https://doi.org/10.1371/journal.pone.0159050
Tisarum R, Samphumphung T, Theerawitaya C, Prommee W, Cha-um S (2018). In vitro photoautotrophic acclimatization, direct transplantation and ex vitro adaptation of rubber tree (Hevea brasiliensis). Plant Cell, Tissue and Organ Culture 133:215-223. https://doi.org/10.1007/s11240-017-1374-5
Uthup TK, Karumamkandathil R, Ravindran M, Saha T (2018). Heterografting induced DNA methylation polymorphisms in Hevea brasiliensis. Planta 248:579-589. https://doi.org/10.1007/s00425-018-2918-6
Venkatachalam P, Priya P, Amma CS, Thulaseedharan A (2004). Identification, cloning and sequence analysis of a dwarf genome-specific RAPD marker in rubber tree [Hevea brasiliensis (Muell.) Arg.]. Plant Cell Reports 23:327-332. https://doi.org/10.1007/s00299-004-0833-8
Wang TD, Huang TD, Huang HS, Hua YW (2017). Origin of secondary somatic embryos and genetic stability of the regenerated plants in Hevea brasiliensis. Journal of Rubber Research 20:101-116. https://doi.org/10.1007/BF03449145
Yang X, Blagodatsky S, Marohn C, Liu H, Golbon R, Xu J, Cadisch G (2019). Climbing the mountain fast but smart: Modelling rubber tree growth and latex yield under climate change. Forest Ecology and Management 439:55-69. https://doi.org/10.1016/j.foreco.2019.02.028
Yao X, Chen X, Wang J, Zhou J, Cai M, Lin W (2017). Effect of clonal rootstocks on the growth and yield of Hevea rubber. Journal of Rubber Research 20:203-212. https://doi.org/10.1007/BF03449152
Zhao H, Jia RZ, Zhu YJ, Guo AP, Zeng HC, Peng M (2015). Efficient induction, proliferation, and regeneration of rubber tree (Hevea brasiliensis Müell. Arg) via somatic embryogenesis. The Journal of Animal and Plant Sciences 25:134-140.
How to Cite
Open Access Journal:
The journal allows the author(s) to retain publishing rights without restriction. Users are allowed to read, download, copy, distribute, print, search, or link to the full texts of the articles, or use them for any other lawful purpose, without asking prior permission from the publisher or the author.