scholarly journals Induksi mutasi Stevia rebaudiana dengan perendaman kolkisin secara in vitro (Induced mutation of Stevia rebaudiana through colchicine soaking in vitro)

2018 ◽  
Vol 86 (1) ◽  
Author(s):  
Masna Maya SINTA ◽  
Ni Made Armini WIENDI ◽  
Syarifah Iis AISYAH

Stevia rebaudiana Bert. is a plant producing steviol glycosides that have 200-300 times sweeter than sucrose. These steviol glycosides are produced in the leaves and then spread to all parts of the plant including stems. The use of superior stevia planting material is important for stevia sugar industry. One of the stevia breeding programme is to increase genetic diversity through colchicine soaking to produce polyploid plants. Polyploid plants usually have higher vigor than diploid plants. The purpose of this research was to induce genetic diversity of stevia through colchicine soaking in vitro. Single nodes of sterile stevia clone BS were soaked in colchicine at the concentration of 0.01; 0.02; 0.04; 0.08 and 0.1% for 48 and 72 hours, and in sterile aquadest as a control. Plantlet subcultures were done until MV4 (mutant vegetative 4). Putative mutants were observed by plantlet vigor and stomata analyses on MV5. Vigor of plantlets was observed by counting the number of leaves, nodes, roots, fresh weight and dry weight of the plantlet. Stomata analysis was performed by calculating stomata density, stomata size and chloroplast number in stomata guard cells. Results showed that colchicine soaking treatment increased significantly fresh weight and dry weight of putative mutants. Colchicine soaking treatment increased chloroplast number on stomata guard cell and stomata size, but decreased stomata density. Stevia soaked in colchicine for 48 hours at concentration 0.01-0.04% produce putative mutants with high chromosome numbers. [Key words: poliploidy, stomata, chloroplast, mutant]AbstrakStevia rebaudiana Bert. merupakan tanaman penghasil glikosida steviol yang memiliki tingkat kemanisan 200-300 kali lebih tinggi dibandingkan sukrosa. Glikosida steviol ini diproduksi di daun yang kemudian disalurkan ke bagian tanaman lainnya termasuk batang. Penggunaan klon terbaik stevia merupakan salah satu kunci penting keberhasilan industri gula stevia. Salah satu program pemuliaan tanaman stevia adalah meningkatkan keragaman tanaman melalui mutasi dengan kolkisin sehingga menghasilkan tanaman poliploid. Tanaman poliploid umumnya memiliki vigor lebih baik dibandingkan tanaman diploid. Tujuan dari penelitian ini adalah untuk meningkatkan keragaman stevia melalui peren-daman kolkisin in vitro. Buku tunggal steril stevia klon BS direndam dalam kolkisin dengan konsentrasi 0,01; 0,02; 0,04; 0,08 dan 0,1% selama 48 dan 72 jam dengan perendaman dalam air steril sebagai kontrol. Sub kultur dilakukan hingga MV4 (mutan vegetatif 4). Pengamatan mutan putatif dilakukan meliputi analisis morfologi dan stomata pada MV5.  Analisis morfologi dilakukan dengan mengamati jumlah daun, buku, akar, bobot basah serta bobot kering planlet. Analisis stomata dilakukan dengan menghitung kerapatan stomata, ukuran stomata serta jumlah kloroplas pada sel penjaga stomata. Hasil menunjukkan bahwa perendaman stevia pada kolkisin meningkatkan bobot basah serta bobot kering stevia in vitro. Perlakuan perendaman kolkisin meningkatkan jumlah kloroplas pada sel penjaga stomata serta ukuran stomata namun menurunkan kerapatan stomata. Perendaman stevia selama 48 jam pada konsentrasi kolkisin 0,01-0,04% menghasilkan mutan putatif dengan jumlah kromosom tertinggi.[Kata kunci: poliploidi, stomata, kloroplas, mutan]

2017 ◽  
Vol 48 (5) ◽  
Author(s):  
Al-Obaidy & Khierallah

This research was conducted to study the effect of some plant growth regulators on in vitro shoots multiplication of stevia (Stevia rebaudiana Bertoni). The experiments included tests of various combinations of KIN with IBA or IAA in the shoot multiplication. Results indicated that KIN at 1.0 mg. L-1 plus 0.3 mg. L-1 of IBA produced the highest number of shoots (3.5 shoots) while KIN at 1.5 mg. L-1 plus IBA at 1.0 mg. L-1 produced the lowest shoot length (1.14 cm).  Hormone free medium produced the highest rate of the leaves number reached 28.56 leaves. KIN and IBA interaction increased fresh and dry weight significantly.   Treatment contained 2.0 mg -1 KIN plus 0.3 mg. L-1 IBA produced the highest fresh weight (1.739 g) while 0.5 mg. L-1 KIN and 0.3 mg. L-1 IBA produced the highest dry weight (0.822 g). As for the effect of interaction between the IAA and KIN it was significant in the number of shoots formed. Interaction between 1.0 mg. L-1 KIN with 0.1 mg. L-1IAA produced the highest number of shoots (3.8 shoots). Shoots length reached 8.10 cm in the media with 0.3 mg. L-1 IAA only. The highest fresh weight (1.267 g) was achieved with the interaction between 1.0 mg. L-1 KIN and 0.3 mg. L-1 IAA while 0.5 mg. L-1IAA without KIN produced the highest dry weight reached 0.138 g.  Shoots multiplication was improved by incorporation of the cytokinin TDZ in culture media. Shoots number, fresh and dry weights were increased significantly by adding 0.05 mg. L-1 of TDZ at present of 0.3 mg. L-1 of IBA giving 6.6 shoots, 0.974 g and 0.144 g respectively while shoots length decreased significantly as media without TDZ produced the highest shoots length reached 9.32 cm. The above results can adopt for the successful in vitro shoot multiplication of Stevia plants. 


2018 ◽  
Vol 2018 ◽  
pp. 1-8
Author(s):  
Juan Pablo Quintal Martínez ◽  
Jorge Carlos Ruiz Ruiz ◽  
Maira Rubí Segura Campos

This study was oriented towards encapsulation of S. rebaudiana extract and the study of its release kinetics. The desired encapsulation was achieved by the ionotropic gelation method using sodium alginate and inulin of polymeric constituents. Characterization of the capsules was performed by micrometric properties, encapsulation efficiency, in vitro extract release analysis, and biological activity of released extract. The in vitro release profiles from different capsules were applied on different kinetic models. The prepared capsules were found spherical in shape with diameters ranging from 2.07 to 2.63 mm, having the encapsulation efficiencies of 43.77% and 56.53% for phenolic compounds and steviol glycosides, respectively. The best-fit model with the highest correlation coefficient was observed in the Ritger–Peppas model, indicating diffusion controlled principle. The release exponent n value obtained from the Korsmeyer–Peppas model varied between 0.2273 and 1.1719, confirming that the mechanism of S. rebaudiana extract bioactive compounds release was diffusion controlled.


Materials ◽  
2021 ◽  
Vol 14 (18) ◽  
pp. 5340
Author(s):  
Alicja Tymoszuk

The interactions between nanoparticles and plant cells are still not sufficiently understood, and studies related to this subject are of scientific and practical importance. Silver nanoparticles (AgNPs) are one of the most commonly produced and used nanomaterials. This study aimed to investigate the influence of AgNPs applied at the concentrations of 0, 50, and 100 mg·L−1 during the process of in vitro germination as well as the biometric and biochemical parameters of developed seedlings in three vegetable species: Solanum lycopersicum L. ‘Poranek’, Raphanus sativus L. var. sativus ‘Ramona’, and Brassica oleracea var. sabellica ‘Nero di Toscana’. The application of AgNPs did not affect the germination efficiency; however, diverse results were reported for the growth and biochemical activity of the seedlings, depending on the species tested and the AgNPs concentration. Tomato seedlings treated with nanoparticles, particularly at 100 mg·L−1, had shorter shoots with lower fresh and dry weights and produced roots with lower fresh weight. Simultaneously, at the biochemical level, a decrease in the content of chlorophylls and carotenoids and an increase in the anthocyanins content and guaiacol peroxidase (GPOX) activity were reported. AgNPs-treated radish plants had shorter shoots of higher fresh and dry weight and longer roots with lower fresh weight. Treatment with 50 mg·L−1 and 100 mg·L−1 resulted in the highest and lowest accumulation of chlorophylls and carotenoids in the leaves, respectively; however, seedlings treated with 100 mg·L−1 produced less anthocyanins and polyphenols and exhibited lower GPOX activity. In kale, AgNPs-derived seedlings had a lower content of chlorophylls, carotenoids, and anthocyanins but higher GPOX activity of and were characterized by higher fresh and dry shoot weights and higher heterogeneous biometric parameters of the roots. The results of these experiments may be of great significance for broadening the scope of knowledge on the influence of AgNPs on plant cells and the micropropagation of the vegetable species. Future studies should be aimed at testing lower or even higher concentrations of AgNPs and other NPs and to evaluate the genetic stability of NPs-treated vegetable crops and their yielding efficiency.


2020 ◽  
Vol 107 (2) ◽  
pp. 171-178
Author(s):  
Aušra Blinstrubienė ◽  
Natalija Burbulis ◽  
Neringa Juškevičiūtė ◽  
Rasa Žūkienė

Author(s):  
M Irshad ◽  
M Idrees ◽  
A Tariq ◽  
ML Pathak ◽  
M Hanif ◽  
...  

The aim of present research was to study the genetic diversity among Asparagus species and its cultivars using morphological characteristics and RAPD markers. In-vitro and field experiments of 14 germplasm sources of Asparagus species and its cultivars were conducted at the Kohat University of Science and Technology to estimate the comparative performance. Highest genotypic variance, phenotypic variance, genotypic and phenotypic coefficient of variance were observed in shoot height in shoot related traits were 318.40, 320.30, 939.34 and 944.96, respectively, whereas highest values for root related traits were observed in root length that were 21.84, 22.35, 141.60 and 144.91 respectively. Among the shoot related traits maximum heritability, genetic advance and genetic gain were calculated for shoot length, while in root related traits these characters were highest for root dry weight. RAPD markers were used to evaluate genetic diversity analysis of 14 germplasm sources of Asparagus species and its cultivars. RAPD markers generated a total of 247 bands, of these 239 bands were polymorphic with average of 34.1 bands per primer. Cluster analysis based on Neighbor Joining methods showed that wild species (A. adsendens, A. densiflorus, A. capitatus, A. gracelus, A. plumosus, A. racemosus and A. setaceus) were genetically distant from A. officinalis and its cultivars (Abril, Apollo, Gersengum, Huchel, Para seletion and Taranga). The results of the present investigations could be particularly used for authentic identification and would be useful for evaluation of genetic improvement of Asparagus species and its cultivars. J. Biodivers. Conserv. Bioresour. Manag. 2019, 5(1): 13-24


2016 ◽  
Vol 27 (2) ◽  
pp. 128-135 ◽  
Author(s):  
J Akte ◽  
S Yasmin ◽  
MJH Bhuiyan ◽  
F Khatun ◽  
J Roy ◽  
...  

Five rice varieties viz. Binadhan-4, Binadhan-5, Binadhan-6, Binadhan-10 and Iratom-24 were evaluated in vitro under different water stress conditions. Several parameters such as germination percentage, shoot length, root length, shoot-root ratio, fresh weight, dry weight, turgid weight, relative water content and proline accumulation were studied. Drought condition was created by MS medium supplemented with five treatments of PEG, with a control such as 0%, 1%, 2%, 3% and 4% of PEG. The highest germination (100%) was found in the variety Binadhan-10 under low water stress conditions induced by 1% PEG. Similarly, the highest percentage of germination was found in all varieties under control condition (0% PEG). The lowest percentage of germination was obtained in the variety Iratom-24. But under severe stress (4% PEG), the highest percentage of germination was found only in the variety Binadhan-10. Moreover, the variety Binadhan-10 was found to be the best at 4% PEG for shoot length, root length, shoot-root ratio, relative water content and also the best at 1% PEG for fresh weight, dry weight, turgid weight. Water stress decreased relative water content and increased proline accumulation in rice. The highest relative water content was recorded in the variety Binadhan-10 and the lowest value recorded in the variety Binadhan-5. The highest proline content was obtained from the binadhan-6 at the highest treatment (4% PEG). Binadhan-10 showed the best performance almost in all the parameters under drought stress because of its own nature of tolerancy.Progressive Agriculture 27 (2): 128-135, 2016


2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Hassan O. Shaikhaldein ◽  
Fahad Al-Qurainy ◽  
Mohammad Nadeem ◽  
Salim Khan ◽  
Mohamed Tarroum ◽  
...  

Abstract Silver nanoparticles (AgNPs) are presently the most commonly generated engineered nanomaterials and are found in a wide range of agro-commercial products. The present study was designed to synthesize AgNPs biologically using Ochradenus arabicus leaves and investigate their effect on the morphophysiological properties of Maerua oblongifolia raised in vitro. Physicochemical methods (ultraviolet–visible spectroscopy, Fourier transform infrared spectroscopy, and transmission electron microscopy were performed for characterization and for obtaining microphotographs of the AgNPs. Shoots of M. oblongifolia (2–3 cm) grown in Murashige and Skoog medium supplemented with different concentrations of AgNPs (0, 10, 20, 30, 40, or 50 mg L−1) were used. Following 6 weeks of in vitro shoot regeneration, the shoot number, shoot length, leaf number, fresh weight, dry weight, chlorophyll content, total protein, proline level, and antioxidant enzyme activities of the plants were quantified. We found that 20 mg L−1 AgNPs increased the shoot number, shoot length, fresh weight, dry weight, and chlorophyll content of the plants. The maximum total protein was recorded in plants that were administered the lowest dose of AgNPs (10 mg L−1), while high concentrations of AgNPs (40 and 50 mg L−1) increased the levels of proline and the enzymes superoxide dismutase and catalase. Our results indicate that green-synthesized AgNPs may be of agricultural and medicinal interest owing to their effects on plants in vitro.


1981 ◽  
Vol 61 (3) ◽  
pp. 665-671 ◽  
Author(s):  
G. LAFOND ◽  
L. E. EVANS

The effect of the "leafless" mutations (in which tendrils replace leaflets, af, and the stipules are reduced to a vestigial form, st) upon chlorophyll content, total protein, soluble protein, ribulose biphosphate carboxylase activity, and fresh weight/dry weight ratio were studied. Comparison of three near-isogenic lines of Century, Freezer and Trapper for the genotypes af af st st (leafless), af af + + (semi-leafless) and + + st st (reduced stipules) were compared with the conventional (+ + + +) plants. Chlorophyll contents were reduced by as much as 50% in the leafless phenotypes, as compared to the leafed plants. The leafless phenotype showed significantly higher levels of total nitrogen than the leafed plants. Tendrils had significantly lower levels of soluble protein than leaflets in two of the three cultivars examined. The fresh weight/dry weight ratio was significantly higher in the leafless phenotypes than the leafed types as a result of changes in morphology, going from a leaflet to a tendril. The internal arrangement of cells in the tendril was typical of pith and cortex tissue with very few intercellular spaces between the parenchyma cells. The intercellular space index within the tendrils was significantly lower than in the leaflet. In vitro measurements of ribulose biphosphate carboxylase activity were calculated per unit fresh weight and per unit chlorophyll. No significant differences were found between the different phenotypes per unit fresh weight; however, when the activity was measured per unit chlorophyll, the values for tendrils were significantly higher than for leaflets.


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