scholarly journals Induksi Tetraploid Tunas Pucuk Jeruk Siam Simadu (Citrus nobilis Lour) Menggunakan Kolkisin secara In Vitro

2015 ◽  
Vol 43 (1) ◽  
pp. 66 ◽  
Author(s):  
Fitri Yulianti ◽  
Agus Purwito ◽  
Ali Husni ◽  
Diny Dinarti

Seedless fruit is one of the criteria (necessary) to improve the quality of Simadu tangerine. The most effective method to obtain seedless triploid cultivars is hybridisation between tetraploid and diploid parents. Simadu tangerine is a diploid plant. Tetraploid Simadu tangerine can be obtained with doubling chromosome using colchicines.The aim of this research was to obtain tetraploid Simadu tangerine shoot which would serve as parent to produced seedless Simadu tangerine. Shoot-tips of Simadu tangerine without leaves were treated with colchicines at four different concentrations (0, 0.1, 0.2, and 0.3%) for 3 hours. The results showed that the high concentration of 0.3% reduced survival rate. The colchicine treatments reduced growth of shoot-tip of Simadu Tangerine.The leaves of colchicines treated shoots were thicker than control. Leaves from control (0% colchicine) and 0.1% colchicine treated shoots had 8.67 and 18.25 chloroplast per pair of guard cells. Compared to those of control, leaves with 0.1% colchicine had lower stomatal density, and larger stomatal size. It appeared that 0.1% colchicine treatment resulted in tetraploid Simadu Tangerine Shoot.<br />Keywords:chloroplasts, doubling chromosomes, stomatal size, stomatal density

2019 ◽  
Vol 28 (1) ◽  
pp. 13
Author(s):  
Dewi Pramanik ◽  
Herni Shintiavira ◽  
Budi Winarto

<p>Anggrek <em>Phalaenopsis</em> memiliki nilai komersial yang tinggi, karena keindahannya dapat dinikmati sepanjang tahun. Hal tersebut berdampak pada kebutuhan benih tanaman yang semakin meningkat. Salah satu cara penyediaan benih secara massal adalah melalui perbanyakan klonal secara in vitro sehingga perlu dilakukan studi kualitas regeneran hasil perbanyakan klonal untuk menjamin ketersediaan benih dengan kualitas baik. Penelitian bertujuan menguji kualitas regeneran yang dihasilkan dari perbanyakan klonal secara in vitro beberapa varietas Phalaenopsis dengan menggunakan eksplan yang berbeda. Penelitian dilaksanakan di Laboratorium Kultur Jaringan, Kebun Percobaan Segunung, Balai Penelitian Tanaman Hias (Balithi) sejak bulan Januari 2014 hingga Mei 2015. Penelitian menggunakan dua faktor, yaitu varietas (Ayu Lestari, Ayu Pratiwi, dan Karindra) dan jenis eksplan (tangkai infloresen, tunas pucuk, dan empulur). Percobaan disusun menggunakan rancangan acak kelompok pola faktorial dan setiap perlakuan diulang tiga kali. Hasil penelitian menunjukan tidak terjadi interaksi yang nyata antara faktor jenis eksplan dan varietas yang diujikan pada semua tahap percobaan. Respon terbaik diperoleh pada eksplan empulur dengan 42,85% eksplan berhasil membentuk kalus pada minggu ke-8 dan hampir 100% kalus tersebut dapat beregenerasi menjadi tunas pada minggu ke-24 dengan tingkat multiplikasi tunas 1,87 kali. Pada minggu ke-32 terbentuk rata-rata 3,13 daun per planlet dengan 2,47 cm panjang daun, 1,36 cm lebar daun, 1,52 akar per planlet, dan panjang akar per planlet mencapai 1,26 cm. Kerapatan stomata memiliki korelasi negatif dengan tingkat abnormalitas planlet. Planlet dengan kerapatan stomata tertinggi dan abnormalitas yang rendah diperoleh pada var. Karindra dan planlet yang berasal dari eksplan empulur dan tunas pucuk. Setelah 8 minggu tahap aklimatisasi, tingkat keberhasilan hidup tertinggi (92%) diperoleh pada tunas yang berasal dari eksplan empulur. Penelitian membuktikan bahwa perbedaan varietas tidak memiliki pengaruh nyata pada tingkat abnormalitas regeneran dan dari eksplan empulur diperoleh jumlah regeneran tertinggi dengan kualitas baik (tingkat abnormalitas rendah).</p><p><strong>Keywords</strong></p><p>Kultur jaringan; Kualitas regeneran; <em>Phalaenopsis</em>; Jenis eksplant</p><p><strong>Abstract</strong></p><p><em>Phalaenopsis</em> orchids have a high commercial value, because of its beauty and it can be enjoyed throughout the year. This condition gives the impact on the increasing demand of the seeds. One of the ways of providing mass seeds is through in vitro clonal propagation. However, it is necessary to study the quality of regenerants of clonal propagation products to ensure the availability of qualified seeds. The aimed of this study was to test the quality of regenerants obtained from in vitro clonal propagation of Phalaenopsis using inflorescence stalk, shoot tips, and pith explants. This research was conducted at Tissue Culture Laboratory, Segunung Experimental Station, Indonesian Ornamental Crops Research Institute (IOCRI) from January 2014 to May 2015. The study used two treatments, varieties (Ayu Lestari, Ayu Pratiwi, and Karindra) and type of explant (inflorescence stalk, shoot tips, and pith). Experiments were prepared using a randomized complete block design with two factors and each treatment was replicated three times. The results showed there were no significant interaction between types of explants and varieties tested in all experiment stages. The best response was obtained using pith explants with 42.85% callus formation in the week eighth and nearly 100% callus can regenerate into shoots at week 24th with the rate of shoot multiplication up to 1.87 times. At week 32th the cultures formed planlets with an average number of leaves of 3.13 and an average size of 2.47 cm x 1.36 cm (length x width) and an average number of roots of 1.52 with average length reached 1.26 cm. Stomatal density has negative correlation with plantlet abnormality rate. Plantlets with the highest stomatal density and low abnormality were obtained in var. Karindra and plantlet derived from explant pith and shoot buds. After 8 weeks of acclimatization stage, the highest survival rate (92%) was obtained on the shoot originating from pith explant. This study proved that varietal differences did not have a significant effect on regenerant abnormalities, and the highest number of regenerant with good quality (low abnormality rate) was obtained from pith explant.</p>


2015 ◽  
Vol 4 (4) ◽  
pp. 208
Author(s):  
Hanifah - Aini ◽  
Mansyurdin - Mansyurdin ◽  
Suwirmen - Suwirmen

The study about PLB induction of wild Vanda sumatrana Schltr. on MS media suplement with BAP and NAA and ploidisation by colchicine treatment was conducted from December 2014 until November 2015 at the Laboratory of Genetics and Cell Biology and Laboratory of Plant Physiology and Tissue Culture, Biology department, Faculty of Mathematic and Natural Science, Andalas University, Padang. The study aimed to 1) knowing the best concentration of 6-Benzyl amino purin (BAP) and α-Naphtalene acetic acid (NAA) for Protocorm Like Bodies (PLB)  induction from shoot tip of V. sumatrana, 2) knowing the PLB response of V. sumatrana to concentrations and soak period of colchicine and 3) find the effective concentrations and soak period of colchicine to induce tetraploid on PLB of V. sumatrana. Shoot tips from in-vitro cultured of V. sumatrana  were subcultured on Murashinge and Skoog (MS) medium supplement with 3 mg/l BAP + 0,5 mg/l NAA, 3 mg/l BAP and 1,5 mg/l BAP. PLB of diploid V. sumatrana from the best treatment were soaked in 0.05% and 0.1% colchicine for 24 and 48 hours respectively in MS liquid medium, as control were set PLB without colchicine treatment. The results showed that MS medium supplemented with 1.5 mg/l BAP was the best formula to induce PLB. The highest percentage of survival rate of PLB and percentage of survived PLB regenerated shoot was obtained from 0.05% colchicine with 24 hours soak period treatment. The effective treatment to induce tetraploid on PLB of V. sumatrana Schltr. was obtained from 0.05% colchicine solution for 24 hours soak period.


2020 ◽  
Vol 2020 ◽  
pp. 1-10 ◽  
Author(s):  
Dingyi Yang ◽  
Yong Jiang ◽  
Yuqing Wang ◽  
Qianqian Lei ◽  
Xin Zhao ◽  
...  

In this study, flavonoids in lemon seeds (FLS) were used to assess its improvement on the oxidative damage of human embryonic kidney 293T cells (HEK 293T cells) induced by H2O2. In vitro experiments showed that the survival rates of HEK 293T cells treated with different flavonoid concentrations (50 μg/mL, 100 μg/mL, and 150 μg/mL) exceeded 95%, indicating no significant toxic effect. Compared with the normal group, H2O2 (0.3 mmol/L) resulted significantly in oxidative stress injury of HEK 293T cells. The survival rate of the damaged cells increased after treatment with flavonoids, and the survival rate of cells treated with a high concentration (150 μg/mL) of flavonoids was 76.2%. Flavonoids also effectively inhibited H2O2-induced apoptosis. At the same time, flavonoid treatment significantly reduced the malondialdehyde content in cells and increased the levels of catalase (CAT), superoxide dismutase (SOD), glutathione (GSH), and glutathione peroxidase (GSH-Px). Quantitative polymerase chain reaction (qPCR) and Western blot analysis also suggested that FLS upregulated mRNA and protein expressions of CAT, SOD (SOD1, SOD2), GSH (GSH1), and GSH-Px in H2O2-induced oxidative damage of HEK 293T cells. The high-performance liquid chromatography analysis demonstrated that FLS contained six compounds, including gallocatechin, caffeic acid, epicatechin, vitexin, quercetin, and hesperidin. FLS were proven to have a good antioxidant capacity in vitro and improve significantly the oxidative damage of HEK 293T cells induced by H2O2. The biological activity value warrants investigation in additional studies.


1992 ◽  
Vol 40 (6) ◽  
pp. 887 ◽  
Author(s):  
SD Hamill ◽  
MK Smith ◽  
WA Dodd

Alternative breeding strategies, based on colchicine-induced autotetraploids, have been proposed as a means of introducing disease resistance into banana breeding programs. This paper describes techniques for the in vitro induction of banana autotetraploids by the use of colchicine on cultured explants. The technique can be readily applied and large numbers of autotetraploids produced. The optimum treatment involved immersing shoot tips in a 0-5% w/v colchicine solution for 2 h under aseptic conditions. Dimethyl sulfoxide (DMSO) was applied with the colchicine treatments to increase cell permeability and so absorption of colchicine, resulting in the optimum treatment unchanged at 0-5% colchicine, but including the addition of 2% v/v DMSO. Of the shoot tips treated over 30% were induced to the autotetraploid level. Methods for in vitro selection of induced tetraploids from treated diploid plantlets were also developed. Tetraploid plants were more robust with thicker pseudostems, roots and broader leaves than diploids and they could be selected on these morphological characteristics. Mean stomatal lengths of diploid banana plants growing in vitro were significantly smaller (16-0 mum) than the tetraploids (26.9 mum) and were used as a more reliable indicator of ploidy than morphological criteria alone. A root tip squash technique using carbol fuchsin was developed for positive confirmation of ploidy change by chromosome counts- Although chimerism and reversion to the diploid form occurred, it was not considered a problem because of the large number of autotetraploids induced. Stable autotetraploids were recovered and established in the field and were characterised by their large, drooping leaves and thick pseudostems. They have retained these characteristics for more than 3 years in the field.


2004 ◽  
Vol 44 (10) ◽  
pp. 1065 ◽  
Author(s):  
M. K. Smith ◽  
S. D. Hamill ◽  
B. J. Gogel ◽  
A. A. Severn-Ellis

Ginger autotetraploids were produced by immersing shoot tips in a 0.5% w/v colchicine, 2% v/v dimethyl sulfoxide solution for 2 h. Stomatal measurements were used as an early indicator of ploidy differences in culture with mean stomata length of tetraploids (49.2 μm) being significantly larger than the diploid (38.8 µm). Of the 500 shoot tips treated, 2% were characterised as stable autotetraploid lines following field evaluation over several seasons. Results were confirmed with flow cytometry and, of the 7 lines evaluated for distinctness and uniformity, 6 were solid tetraploid mutants and 1 was a periclinal chimera. Significant differences were noted between individual tetraploid lines in terms of shoot length, leaf length, leaf width, size of rhizome sections (knob weight) and fibre content. The solid autotetraploid lines had significantly wider, greener leaves than the diploids, they had significantly fewer but thicker shoots and, although ‘Queensland’ (the diploid parent from which the tetraploids were derived) had a greater total rhizome mass at harvest, its knob size was significantly smaller. From the autotetraploid lines, one line was selected for commercial release as ‘Buderim Gold’. It compared the most favourably with ‘Queensland’ in terms of the aroma/flavour profile and fibre content at early harvest, and had consistently good rhizome yield. More importantly it produced large rhizome sections, resulting in a higher recovery of premium grade confectionery ginger and a more attractive fresh market product.


2021 ◽  
Vol 306 ◽  
pp. 01029
Author(s):  
Farida Yulianti ◽  
Dita Agisimanto

Pontianak tangerine citrus (Siam Pontianak) is the most popular tangerine citrus in Indonesia. Induced mutation with colchicine treatment on Pontianak tangerine was intended to produce polyploid plants. The study aimed to analyze the cytological diversity in colchiploid of Pontianak tangerine citrus. The study was conducted at ICISFRI from January – May 2018. The cytological analysis was performed on a five-year-old of 36 colchiploids of Pontianak tangerine and a mother plant as control. The results showed that the colchiploid plants had higher chromosome numbers, stomatal sizes, and chloroplast numbers than the control. However, the stomatal density in several genotypes did not differ from that of the control. Colchicine treatment resulted in an aneuploid plant with chromosome number 18+x. Based on the clustering analysis results, all colchiploid plants differed from the control with a variability coefficient of 0.65. The colchiploid plants were not grouped according to colchicine concentration and duration of treatment. The increase of colchicine concentration and treatment duration did not correlate with the increasing ploidy number of colchiploid plants. The implication of the research result was the stomatal density, stomatal size, and chloroplast number were useful tools for rapid pre-screening of plant polyploidy.


1970 ◽  
Vol 39 (1) ◽  
pp. 47-50 ◽  
Author(s):  
AKM Sayeed Hassan ◽  
Rahima Khatun

Shoot tips and nodal explants from in vitro growing seedlings of Ficus glomerata Roxb. (Moraceae). showed best shoot induction (88%) on MS medium supplemented with 0.5 mg/l BAP, where maximum number of shoots were produced per culture. In vitro raised shoots rooted well on half strength of MS medium with 2.0 mg/l IBA + 0.1 mg/l NAA. The survival rate of regenerated plantlets was 82%. Key words: Ficus glomerata Roxb.; Shoot proliferation; Micropropagation; Acclimatization DOI: 10.3329/bjb.v39i1.5525Bangladesh J. Bot. 39(1): 47-50, 2010 (June)


1970 ◽  
Vol 18 ◽  
pp. 140-145 ◽  
Author(s):  
Md Abu Hena Mostofa Jamal ◽  
ANM Rubaiyath-Bin Rahman ◽  
Dipak Kumar Paul ◽  
Md Rezuanul Islam

Context: It is necessary to focus on the importance of adopting micropropagation technique for mass propagation of the plantlets in commercial scale as well as conservation and distribution of germplasm. Objective: The present investigation has been designed with a view to establishing protocol of in vitro regeneration of medicinal plant species i,e., Vitex nigundo L (Verbenaceae). Materials and Methods: Shoot tips and nodal segments were used for multiple shoot induction. All explants were cultured on MS medium supplemented with various plant growth regulators. HgCl2 was used as surface sterilizing agent. For in vitro rooting, individual shoots (3-4 cm) were cut from the proliferated shoot cultures and implanted on half and full strength of MS with different concentrations and combinations of NAA and IAA. The cultures were incubated for 16 h photoperiod at 25 ± 2ºC under a fluorescent light. Visual observation of culture was made every week. Data on shoot induction and proliferation and root induction were recorded after three weeks of inoculation and used for calculation. For each treatment 15 explants were used and all the treatments were repeated thrice. Established plantlets were transplanted in earthen pots under natural conditions and the survival rate was recorded. Results: The most effective surface sterilization treatment has been found 0.1 % HgCl2 for 7 minutes. Highest number of shoot was observed in MS medium supplemented with 3.0 mg/ BAP. It was rooted well in full MS containing 2.0 mg/l IAA. The survival rate was 85 % and propagated plantlets were successfully acclimatized in soil. Conclusion: It was observed that shoot tips are more responsive for micropropagation of Vitex nigundo L . Thus the fruitful utilization of rapid clonal propagation, germplasm conservation and distribution of Vitex nigundo, important medicinal plant of Bangladesh, is possible. Keywords: Vitex nigundo; Medicinal plant; Shoot induction; Micropropagation; Regeneration. DOI: http://dx.doi.org/10.3329/jbs.v18i0.8790 JBS 2010; 18(0): 140-145


2013 ◽  
Vol 25 (2) ◽  
pp. 133-140 ◽  
Author(s):  
Małgorzata Zalewska ◽  
Dariusz Kulus

ABSTRACT Chrysanthemums are amongst the most economically important flowers in the world. The protection and storage of these valuable genetic resources is of great importance. Today, cryopreservation, or the storage of biological material at the temperature of liquid nitrogen (-196°C), is believed to be the most promising long-term storage method. To optimise the cryopreservation protocol, the shoot tips of Chrysanthemum × grandiflorum /Ramat./ Kitam. ‘Lady Orange’ and ‘Lady Salmon’ mutants were cryopreserved using the encapsulation-dehydration technique. During the experiment, the influence of sucrose concentration (2, 3 and 6%) during preculture and the concentration of kinetin (0.25, 0.5, 0.75 and 1.0 mg dm-3) in the regrowth medium were tested. A higher survival rate was observed for ‘Lady Salmon’. In general, the media with higher sucrose levels provided the best survival and recovery rates (35-40%). Kinetin had no influence on the survival rate; however, it influenced the morphogenesis of the plants. The lowest number of explants forming multiple shoots was observed on the medium with the lowest sucrose (during preculture) and kinetin (in the recovery medium) concentration. On the other hand, the best rhizogenesis efficiency was observed when 0.25 mg dm-3 kinetin was added. In conclusion, the composition of both preculture and recovery media need to be adjusted to single cultivars. The use of 3% sucrose (preculture) and 0.25 mg dm-3 kinetin (recovery) seems reasonable, since it guarantees a satisfying recovery rate of the explants and at the same time prevents the formation of callus and multiple shoots, stimulating the rooting instead.


2014 ◽  
Vol 2014 ◽  
pp. 1-11 ◽  
Author(s):  
L. C. Cruz ◽  
J. E. S. Batista ◽  
A. P. P. Zemolin ◽  
M. E. M. Nunes ◽  
D. B. Lippert ◽  
...  

We characterized, for the first time, the quality and identity of Brazilian Pampa biome honey and its antioxidant propertiesin vitro(FRAP, DDPH and ABTS). The potential protective effect of honey against oxidative stress induced by iron (Fe) and paraquat, (PQ) in aDrosophila melanogastermodel (in vivo) was also tested. The results indicated that all honey samples tested showed antioxidant activityin vitro. Flies treated with honey showed increased lifespan and were protected against oxidative stress induced by Fe and PQ. Despite the high concentration of sugars in honey (approximately 70–80%), our results demonstrate a hypoglycemic-like effect of honey inDrosophila. Thus, this study demonstrates the high quality of Brazilian Pampa biome honey as well as its significant antioxidant activityin vitroandin vivo, pointing to the potential use of this natural product as an alternative in the therapy of oxidative stress-associated diseases.


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