scholarly journals SUSTAINABLE MICROPROPAGATION OF SELECTED Stevia rebaudiana Bertoni GENOTYPES

2019 ◽  
Vol 18 (6) ◽  
pp. 47-56
Author(s):  
Begum Kaplan ◽  
Selda Duraklioglu ◽  
Kenan Turgut

Stevia rebaudiana Bertoni is a perennial plant belonging to Asteraceae family and its leaves contain steviol glycosides (SGs) that are 150 to 300 times sweeter than sucrose. The sweeteners obtained from S. rebaudiana can be safely used by diabetics as insulin secretion is not required during digestion of this sweetener. As it has zero calories, it is also used in diet products. Adaptation studies for Stevia conducted in Antalya, Turkey have shown that the stevia plant could easily be cultivated as a perennial. However, the lack of a sustainable vegetative propagation method creates a significant problem for stevia production. In the generatively populations, homogeneity and therefore quality are decreased because of cross-pollination. Stevia, as a self-incompatible and cross-pollinated species, has been shown to have very high genetic diversity. Therefore, development of a sustainable in vitro propagation method to prevent genetic heterogeneity of selected varieties is crucial for stevia cultivation. The aim of this study was to evaluate 2 different gelling agents (plant agar and Gelrite) and 20 different growth regulators combinations. The results demonstrated an approximately 200-fold multiplication rate obtained within 13 weeks using MS medium supplemented with 0.5 mg·dm–3 BAP and 0.25 mg·dm–3 kinetin and solidified with Gelrite. Average stevioside and rebaudioside A contents in in vitro propagated plant samples were found to be 8.1% and 8.6%, respectively.

2016 ◽  
Vol 79 (2) ◽  
Author(s):  
. SUMARYONO ◽  
Masna Maya SINTA

AbstractStevia (Stevia rebaudiana Bertoni) is a natural zero-calorie sweetener plant grown in a high population density.Tissue culture technique is useful for rapid mass propagationof plants to provide superior planting materials. Experimentswere conducted to increase growth and multiplication ofshoots and vigor of plantlets of stevia. Explants used wereapical and axillary buds from plantlets grown on MS mediumwithout plant growth regulators. Combinations of BA andIAA at different concentrations were used for shoot growthand multiplication, whereas plant growth retardants(ancymidol and paclobutrazol) and light intensity were usedfor plantlet vigor. The results showed that stevia explantscultured on MS medium without plant growth regulatorsproduced the highest shoots (4.5 cm) with two shoots perexplant. The best multiplication rate of shoots were found onMS medium added with 1.13 mg/L BA combined with0.35 mg/L IAA which produced on average 4.5 shoots and11.9 nodes per initial explant. Ancymidol and paclobutrazolconcentrations affected significantly growth and vigor ofstevia plantlets. Increasing the concentration of ancymidoland paclobutrazol decreased plantlet height and biomassfresh weight, but increased stem diameter. Paclobutrazol at0.1 mg/L was the best treatment to increase the vigor ofstevia plantlets. Light intensity at 20 µmol/m 2 /s gave betterplantlet vigor than other light intensities. It can be concludedthat multiplication of stevia shoots should be grown on MSmedium supplemented with 1.13 mg/L BA + 0.35 mg/L IAAand the vigor of the shoots can be increased by culturing onMS medium containing 0.1 mg/L paclobutrazol underfluorescence lamps with 20 µmol/m 2 /s light intensity.AbstrakStevia (Stevia rebaudiana Bertoni) adalah tanamanpemanis alami nir-kalori yang ditanam dengan kerapatanpopulasi yang sangat tinggi. Teknik kultur jaringan dapatdigunakan untuk perbanyakan tanaman secara massal dancepat untuk menyediakan bahan tanam unggul. Penelitiantelah dilakukan untuk meningkatkan pertumbuhan danmultiplikasi tunas dan keragaan planlet stevia. Eksplan yangdigunakan adalah tunas pucuk dan tunas samping dari planletyang ditumbuhkan pada medium MS tanpa zat pengaturtumbuh. Kombinasi BA dan IAA dengan konsentrasi yangberbeda digunakan untuk pertumbuhan dan multiplikasitunas, sedangkan zat penghambat tumbuh (ansimidol danpaklobutrazol) serta intensitas cahaya digunakan untukkeragaan planlet. Hasil penelitian menunjukkan bahwaeksplan stevia yang ditumbuhkan pada medium MS tanpa zatpengatur tumbuh menghasilkan tunas paling tinggi (4,5 cm)dengan dua tunas per eksplan. Multiplikasi tunas terbaikdiperoleh pada medium dengan BA 1,13 mg/L yangdikombinasikan dengan IAA 0,35 mg/L yang menghasilkan4,5 tunas dan 11,9 ruas per eksplan awal. Konsentrasiansimidol dan paklobutrazol berpengaruh nyata terhadappertumbuhan dan keragaan planlet stevia. Meningkatnyakonsentrasi ansimidol dan paklobutrazol menurunkan tinggiplanlet dan bobot basah biomassa, tetapi meningkatkandiameter batang. Paklobutrazol pada konsentrasi 0,1 mg/Lmerupakan perlakuan terbaik untuk meningkatkan keragaanplanlet stevia. Intensitas cahaya pada 20 µmol/m 2 /detikmemberikan keragaan planlet yang lebih baik dibandingkanintensitas cahaya yang lain. Dapat disimpulkan bahwamultiplikasi tunas stevia sebaiknya dilakukan pada mediumMS ditambah BA 1,13 mg/L + IAA 0,35 mg/L dan keragaanplanlet dapat ditingkatkan dengan menanam planlet padamedium MS ditambah paklobutrazol 0,1 mg/L di bawahlampu fluoresen dengan intensitas cahaya 20 µmol/m 2 /detik.


2019 ◽  
Vol 38 (4) ◽  
pp. 1341-1353 ◽  
Author(s):  
Simone Ribeiro Lucho ◽  
Marcelo Nogueira do Amaral ◽  
Priscila Ariane Auler ◽  
Valmor João Bianchi ◽  
María Ángeles Ferrer ◽  
...  

2015 ◽  
Vol 61 (1) ◽  
pp. 50-63 ◽  
Author(s):  
Aleksandra Luwańska ◽  
Aleksandra Perz ◽  
Grażyna Mańkowska ◽  
Karolina Wielgus

Summary Stevia is a plant attracting attention due to its capability to synthesize a group of chemical compounds with sweet taste, i.e. steviol glycosides. Steviol glycosides are successfully applied as a natural sweetener, and some of them have also therapeutic properties. This paper presents available information on the use of stevia plant tissue cultures with the focus on their potential application in food industry. Detailed analysis was done concerning the research employing in vitro culture techniques and the use of them in biosynthesis of secondary metabolites of high importance for the food industry. Both established achievements and most recent publications on stevia were used for assessment of practical applications of the aforementioned techniques and prospects for their development.


2013 ◽  
Vol 59 (3) ◽  
pp. 251-255 ◽  
Author(s):  
Gabriela Kunová ◽  
Vojtěch Rada ◽  
Adrien Vidaillac ◽  
Ivana Lisova

2011 ◽  
Vol 91 (1) ◽  
pp. 1-27 ◽  
Author(s):  
Ashok Kumar Yadav ◽  
S. Singh ◽  
D. Dhyani ◽  
P. S. Ahuja

Yadav, A. K., Singh, S., Dhyani, D. and Ahuja, P. S. 2011. A review on the improvement of Stevia [Stevia rebaudiana (Bertoni)]. Can. J. Plant Sci. 91: 1–27. Stevia rebaudiana (Bertoni) is a herbaceous perennial plant (2n=22) of genus Stevia Cav., which consists of approximately 230 species of herbaceous, shrub and sub-shrub plants. Leaves of stevia produce diterpene glycosides (stevioside and rebaudiosides), non-nutritive, non-toxic, high-potency sweeteners and may substitute sucrose as well as other synthetic sweetners, being 300 times sweeter than sucrose. In addition to its sweetening property, it has medicinal values and uses. Stevia is self-incompatible plant and the pollination behaviour is entomophilous. Rebaudioside-A is of particular interest among the glycosides produced in the leaves of stevia because of the most desirable flavour profile, while, stevioside is responsible for aftertaste bitterness. Development of new varieties of S. rebaudiana with a higher content of rebaudioside-A and a reduced content of stevioside is the primary aim of plant breeders concerned with the improvement and utilization of this source of natural sweeteners. The proportions of rebaudioside-A and -C are controlled by a single additive gene known to be co-segregating suggesting synthesis by the same enzyme. Stevioside and rebaudioside-A are negatively correlated, while rebaudioside-A and -C are positively correlated. Conventional plant breeding approaches such as selection and intercrossing among various desirable genotypes is the best method for improving quality traits in a highly cross-pollinated crop like stevia. Various plant types with larger amounts of specific glycoside have already been patented, such as RSIT 94-1306, RSIT 94-75, RSIT 95-166-1 through selection and intercrossing. Composites and synthetics can be used to capture part of the available heterosis because of the high degree of natural out-crossing and the absence of an efficient system of pollination control. Synthetics and composites like “AC Black Bird” and “PTA-444” have already been developed. Polyploidy results in better adaptability of individuals and increased organ and cell sizes. Tetraploids have larger leaf size, thickness and have potential use in increasing biomass and yield in comparison with diploid strains. Characters of interest with low variability in the population may be improved through mutation breeding. Use of biotechnological approaches, such as tissue culture for the mass propagation of elite genotypes, anther culture for development of pure homozygous doubled haploid and molecular marker technology for identification of marker loci linked to rebaudioside-A trait, can create new opportunities for plant breeders. Understanding the mechanism and pathway of biosynthesis of steviol glycosides can help to improve the glycoside profile by up-regulation and down-regulation of genes.


2016 ◽  
Vol 79 (2) ◽  
Author(s):  
. SUMARYONO ◽  
Masna Maya SINTA

AbstractStevia (Stevia rebaudiana Bertoni) is a natural zero-calorie sweetener plant grown in a high population density.Tissue culture technique is useful for rapid mass propagationof plants to provide superior planting materials. Experimentswere conducted to increase growth and multiplication ofshoots and vigor of plantlets of stevia. Explants used wereapical and axillary buds from plantlets grown on MS mediumwithout plant growth regulators. Combinations of BA andIAA at different concentrations were used for shoot growthand multiplication, whereas plant growth retardants(ancymidol and paclobutrazol) and light intensity were usedfor plantlet vigor. The results showed that stevia explantscultured on MS medium without plant growth regulatorsproduced the highest shoots (4.5 cm) with two shoots perexplant. The best multiplication rate of shoots were found onMS medium added with 1.13 mg/L BA combined with0.35 mg/L IAA which produced on average 4.5 shoots and11.9 nodes per initial explant. Ancymidol and paclobutrazolconcentrations affected significantly growth and vigor ofstevia plantlets. Increasing the concentration of ancymidoland paclobutrazol decreased plantlet height and biomassfresh weight, but increased stem diameter. Paclobutrazol at0.1 mg/L was the best treatment to increase the vigor ofstevia plantlets. Light intensity at 20 µmol/m 2 /s gave betterplantlet vigor than other light intensities. It can be concludedthat multiplication of stevia shoots should be grown on MSmedium supplemented with 1.13 mg/L BA + 0.35 mg/L IAAand the vigor of the shoots can be increased by culturing onMS medium containing 0.1 mg/L paclobutrazol underfluorescence lamps with 20 µmol/m 2 /s light intensity.AbstrakStevia (Stevia rebaudiana Bertoni) adalah tanamanpemanis alami nir-kalori yang ditanam dengan kerapatanpopulasi yang sangat tinggi. Teknik kultur jaringan dapatdigunakan untuk perbanyakan tanaman secara massal dancepat untuk menyediakan bahan tanam unggul. Penelitiantelah dilakukan untuk meningkatkan pertumbuhan danmultiplikasi tunas dan keragaan planlet stevia. Eksplan yangdigunakan adalah tunas pucuk dan tunas samping dari planletyang ditumbuhkan pada medium MS tanpa zat pengaturtumbuh. Kombinasi BA dan IAA dengan konsentrasi yangberbeda digunakan untuk pertumbuhan dan multiplikasitunas, sedangkan zat penghambat tumbuh (ansimidol danpaklobutrazol) serta intensitas cahaya digunakan untukkeragaan planlet. Hasil penelitian menunjukkan bahwaeksplan stevia yang ditumbuhkan pada medium MS tanpa zatpengatur tumbuh menghasilkan tunas paling tinggi (4,5 cm)dengan dua tunas per eksplan. Multiplikasi tunas terbaikdiperoleh pada medium dengan BA 1,13 mg/L yangdikombinasikan dengan IAA 0,35 mg/L yang menghasilkan4,5 tunas dan 11,9 ruas per eksplan awal. Konsentrasiansimidol dan paklobutrazol berpengaruh nyata terhadappertumbuhan dan keragaan planlet stevia. Meningkatnyakonsentrasi ansimidol dan paklobutrazol menurunkan tinggiplanlet dan bobot basah biomassa, tetapi meningkatkandiameter batang. Paklobutrazol pada konsentrasi 0,1 mg/Lmerupakan perlakuan terbaik untuk meningkatkan keragaanplanlet stevia. Intensitas cahaya pada 20 µmol/m 2 /detikmemberikan keragaan planlet yang lebih baik dibandingkanintensitas cahaya yang lain. Dapat disimpulkan bahwamultiplikasi tunas stevia sebaiknya dilakukan pada mediumMS ditambah BA 1,13 mg/L + IAA 0,35 mg/L dan keragaanplanlet dapat ditingkatkan dengan menanam planlet padamedium MS ditambah paklobutrazol 0,1 mg/L di bawahlampu fluoresen dengan intensitas cahaya 20 µmol/m 2 /detik.


Author(s):  
Miao Si Chiew ◽  
Kok Song Lai ◽  
Sobri Hussein ◽  
Janna Ong Abdullah

Stevia rebaudiana Bertoni from Asteraceae family is commercially valuable for its steviol glycosides (SGs) contents, which is 300 times sweeter than commercial sugar. The bottleneck in Malaysia is the lack of suitable stevia varieties that are able to thrive well under her climatic conditions and still produce high SGs. Mutation induction including gamma irradiation is effective in generating genetic variations and developing new plant varieties with desired traits. This study was aimed to determine the effects of acute gamma irradiation on phenotypic changes and enhancement of SGs contents of Stevia rebaudiana Bertoni variety AKH L1 (herein after will be designated as AKH L1). In vitro shoot tip explants of AKH L1 were subjected to a gamma doses regime of 10Gy to 50Gy, following which phenotypic changes of the irradiated explants and subsequent regenerated plantlets were observed. All irradiated explants exhibited different survival rates, with the lowest at 9.33±8.33% when subjected to 50Gy, while all the control (non-irradiated explants) survived. The LD50 was found to be at 23Gy. Subsequent irradiation of 900 shoot tip explants at 23Gy, produced 468 surviving shoot tips, which were all capable to develop and successfully sub-cultured until the fourth generation, M4. These M4 in vitro mutant plantlets exhibited significant increase in the numbers of leaf (16.07±5.19) and average leaf size (1.12±0.26cm x 0.54±0.15cm). HPLC analysis performed in parallel further revealed the mutant plants contained higher concentrations of stevioside (387.04ppm), rebaudioside A (670.18ppm) and rebaudioside D (106.26ppm) compared to the non-irradiated plantlets, which exhibited 96.87, 194.42 and 28.25ppm, respectively.


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