scholarly journals PENINGKATAN KERAGAMAN GENETIK PURWOCENG MELALUI IRADIASI SINAR GAMMA DAN SELEKSI IN VITRO

2020 ◽  
Vol 19 (2) ◽  
pp. 88
Author(s):  
IKA ROOSTIKA ◽  
IRENG DARWATI ◽  
YUDIWANTI YUDIWANTI
Keyword(s):  
Genetic Variation ◽  
High Temperature ◽  
Gamma Irradiation ◽  
Somatic Embryos ◽  
In Vitro Selection ◽  
Basal Medium ◽  
Root Induction ◽  
Embryogenic Calli ◽  
Low Altitude

<p>ABSTRAK<br />Peningkatkan keragaman genetik purwoceng memerlukan aplikasi<br />teknologi alternatif yang mampu membentuk keragaman baru. Tujuan<br />penelitian adalah untuk meningkatkan keragaman genetik dan toleransi<br />purwoceng terhadap cekaman suhu tinggi melalui iradiasi dan seleksi in<br />vitro. Tahapan penelitian meliputi induksi mutasi kalus embriogenik<br />dengan sinar gamma, seleksi in vitro dengan cekaman suhu tinggi, induksi<br />perakaran  somaklon  putatif,  analisis  keragaman  genetik  secara<br />flowcytometry, dan aklimatisasi somaklon putatif. Iradiasi dilakukan pada<br />dosis 0, 1, 2, 3, 4, dan 5 Krad sedangkan seleksi in vitro dilakukan pada<br />tiga level suhu (20, 25, dan 30 0 C). Induksi perakaran dilakukan dalam dua<br />tahap, dengan menggunakan media DKW atau MS yang mengandung<br />sukrosa 3-6% dengan penambahan IBA atau NAA taraf 0,5-1,5 ppm. Hasil<br />penelitian menunjukkan bahwa kalus purwoceng mampu bertahan hidup<br />pada dosis iradiasi tertinggi (5 Krad). Meningkatnya dosis iradiasi<br />cenderung meningkatkan pendewasaan embrio somatik. Pada tahap seleksi<br />in vitro, kalus purwoceng mampu tumbuh pada kondisi suhu tertinggi<br />(30 0 C). Tingkat proliferasi kalus yang tinggi dan jumlah embrio somatik<br />terbanyak diperoleh dari perlakuan suhu 25 0 C. Embrio somatik yang<br />terbentuk dari perlakuan suhu tinggi tersebut merupakan kandidat<br />somaklon yang toleran suhu tinggi pada lingkungan dataran rendah.<br />Diantara embrio somatik yang terbentuk, hanya embrio yang berasal dari<br />perlakuan suhu 20 0 C saja yang berhasil membentuk planlet. Media yang<br />terbaik untuk induksi perakaran adalah media MS yang mengandung<br />sukrosa 4% dengan penambahan NAA 1,5 ppm. Analisis ploidi pada daun<br />embrio somatik menunjukkan terbentuknya varian yang bersifat tetraploid<br />(4x).<br />Kata kunci: Pimpinella pruatjan, iradiasi sinar gamma, seleksi in vitro,<br />keragaman genetik, suhu tinggi</p><p>ABSTRACT<br />To improve new pruatjan genetic variations, the alternative<br />technology should be applied. The objective of the research was to increase<br />pruatjan genetic variation and tolerance to the high temperature through<br />induced mutation and in vitro selection. The steps of this study were induced<br />mutation of embryogenic callus by gamma irradiation, in vitro selection, root<br />induction of putative somaclones, genetic variation analysis by flowcytometer,<br />and putative somaclones acclimatization. The dosages of gamma irradiation<br />were 0, 1, 2, 3, 4, and 5 Krad. In vitro selection was conducted at three<br />temperatures (20, 25, and 30 0 C). The root induction was conducted in two<br />steps by using DKW or MS media containing of 3-6% sucrose with<br />addition of 0.5-1.5 ppm IBA or NAA. The result showed that embryogenic<br />calli could survive after treatment of the highest gamma irradiation dose. It<br />tends to increase the maturation of somatic embryos. During in vitro<br />selection, embryogenic calli could grow at the highest temperature but the<br />highest callus proliferation and the number of somatic embryos were<br />obtained from 25 0 C. The somatic embryos survived and grew at the high<br />temperature are assumed as somaclones which considered as the<br />candidates of tolerant plants to high temperature that can be developed in<br />the of low altitude area. Among the regenerated somatic embryos, only the<br />20 0 C-derived embryos were successfully form plantlets. The best medium<br />for root induction was MS basal medium containing of 4% sucrose<br />supplemented with 1.5 ppm NAA. The ploidy analysis of somatic embryos<br />leaf showed a tetraploid (4x) variant.<br />Key words: Pimpinella pruatjan, gamma irradiation, in vitro selection,<br />genetic variation, high temperature</p>

scholarly journals Peningkatan Toleransi Alumunium pada Jeruk Batang Bawah dengan Teknik Seleksi In Vitro Berulang

2016 ◽  
Vol 6 (1) ◽  
pp. 33
Author(s):  
Mia Toruan Kosmiatin ◽  
Rosa Yunita ◽  
Ali Husni
Keyword(s):  
In Vitro Selection ◽  
Acid Soil ◽  
Phase 1 ◽  
Aluminum Tolerance ◽  
Basal Medium ◽  
Culture Technique ◽  
Low Ph ◽  
Embryogenic Calli ◽  
Marginal Soil

<p>Aluminum Tolerance Improvement of Rootstock Citrus<br />through Repeated In Vitro Selection. Mia Kosmiatin,<br />Rosa Yunita, and Ali Husni. National orange productivity<br />was trend to decrease because of pathogen attack and<br />reducing of planting area. One of alternative ways to<br />preserve and increase orange productivity was using<br />marginal soil mainly acid soil. This matter pushed the<br />breeder to prepare tolerant rootstock and stable in the acid<br />soil. In vitro culture technique was effective and efficient<br />methods to produce tolerant and stable rootstock in acid soil<br />through simulation of acid soil with addition of high<br />aluminum and low pH in the medium. By the simulation the<br />selection could be done in cell level, so cell was selected<br />after induction of variation. A rootstock which high<br />compatibility with scion, useful rooting, and aluminum<br />tolerance could be increased orange productivity through<br />acid soil development. The research was conducted in 3<br />phase: (1) induction of embryogenic calli, (2) improvement<br />of genetic variation through mutation, and (3) In vitro<br />selection with AlCl3.6H2O for aluminum and low pH tolerant.<br />Immature embryos of rootstock were use as explant. The<br />result showed that the best embryogenic calli were induced<br />on MS basal medium with MW vitamin + NAA 7,5 mg/l +<br />kinetin 0,5 mg/l. Before selection, 1.000 rad dosage was the<br />most tolerant dosage to growth embryogenic calli. After<br />selection, 2.000 rad dosage was the best dosage to produce<br />shoots which stable tolerant to aluminum. Selected 88<br />mutant shoots were produced after three times selection on<br />the same medium which AlCl3.6H2O added at low pH.</p>

scholarly journals Plant Regeneration of Allium victorialis var. platyphyllum Makino via Organogenesis and Somatic Embryogenesis

HortScience ◽  
1996 ◽  
Vol 31 (4) ◽  
pp. 628a-628
Author(s):  
Hak-Tae Lim ◽  
Eun-Ae Lee ◽  
Won-Bae Kim
Keyword(s):  
Somatic Embryogenesis ◽  
Somatic Embryos ◽  
Multiple Shoots ◽  
Root Induction ◽  
Ms Medium ◽  
Mass Propagation ◽  
Embryogenic Calli ◽  
Wild Vegetable ◽  
Shoot Tip Explants

This study was conducted to investigate the possibility of obtaining plantlets via somatic embryogenesis and organogenesis as means of in vitro mass propagation in Allium victorialis var. platyphyllum Makino, one of the most popular wild vegetable plants in Korea. Shoots formed directly when bulb explants of A. victorialis were cultured on MS medium containing 0.2 mg·L–1 NAA and 2.0 mg·L–1 zeatin under 16 hours (light)/8 hours (dark) illumination. The use of leaf and shoot tip explants was not successful, largely due to explant senescence in the present of plant growth regulators. Embryogenic calli were obtained from the bulb explants of A. victorialis on MS medium supplemented with 0.2 mg·L–1 NAA, 0.2 mg·L–1 BAP, and 1.0 mg·L–1 picloram after 4–5 weeks of culture in the dark at 27°C. Upon transfer to shoot-induced MS medium containing 0.2 mg·L–1 NAA and 2.0 mg·L–1 zeatin, embryogenic calli gave rise to numerous somatic embryos, which subsequently developed into multiple shoots after 3 months of culture under 16 hours(light)/8 hours (dark) illumination. For root induction, regenerated shoots were transferred to MS medium added with 1.0 mg·L–1 NAA. Regenerants with well-developed roots were potted in an artificial soil mixture of vermiculite (1) and perlite (1).

Somatic Embryogenesis and In vitro Plant Regeneration in Vigna trilobata (L.) Verd. - A Potential Pasture Legume

1970 ◽  
Vol 19 (1) ◽  
pp. 89-99
Author(s):  
K. Choudhary ◽  
M. Singh ◽  
M. S. Rathore ◽  
N. S. Shekhawat
Keyword(s):  
Somatic Embryogenesis ◽  
Growth Regulators ◽  
Somatic Embryos ◽  
Basal Medium ◽  
Long Term Study ◽  
Continuous Application ◽  
First Time ◽  
Pasture Legume

This long term study demonstrates for the first time that it is possible to propagate embryogenic Vigna trilobata and to subsequently initiate the differentiation of embryos into complete plantlets. Initiation of callus was possible on 2,4-D. Somatic embryos differentiated on modified MS basal nutrient medium with 1.0 mg/l  of 2,4-D and 0.5 mg/l  of Kn. Sustained cell division resulted in globular and heart shape stages of somatic embryos. Transfer of embryos on to a fresh modified MS basal medium with 0.5 mg/l of Kn and 0.5 mg/l of GA3 helped them to attain maturation and germination. However, the propagation of cells, as well as the differentiation of embryos, were inhibited by a continuous application of these growth regulators. For this reason, a long period on medium lacking these growth regulators was necessary before the differentiation of embryos occurred again. The consequences for improving the propagation of embryogenic cultures in Vigna species are discussed. Key words: Pasture  legume, Vigna trilobata, Globular, Heart shape, somatic embryogenesis D.O.I. 10.3329/ptcb.v19i1.4990 Plant Tissue Cult. & Biotech. 19(1): 89-99, 2009 (June)

scholarly journals In Vitro Micropropagation of the Medicinal Plant Physalis angulata L.

2016 ◽  
Vol 8 (2) ◽  
pp. 161-163
Author(s):  
Owk ANIEL KUMAR ◽  
Songa RAMESH ◽  
Sape SUBBA TATA
Keyword(s):  
Large Scale ◽  
Leaf Disc ◽  
Basal Medium ◽  
Shoot Proliferation ◽  
Shoot Multiplication ◽  
Medicinal Herb ◽  
Root Induction ◽  
Survival Percentage ◽  
Physalis Angulata

Physalis angulata L. is an important medicinal herb. An efficient direct adventitious plant regeneration protocol was developed for large scale propagation using leaf disc as explants. The explants were cultured on MS basal medium supplemented with 0.25-3.0 mg/L 6-benzyl amino purine (BAP) for primary shoot proliferation. Inclusion of indole-3-acetic acid (IAA) and gibberellic acid (GA3) in the culture medium along with BAP promoted a higher rate of shoot multiplication. The maximum number of shoots was produced in MS + BAP (1.0 mg/L) + IAA (0.5 mg/L) + GA3 (0.20 mg/L) after the third subculture. An average of 152.8 ± 0.40 shoots were produced from each leaf disc. For root induction the shootlets were transferred to MS medium supplemented with different concentrations of indole-3-butyric acid (IBA). The highest percentage of root induction was observed in 1.0 mg/L (IBA). Rooted plants were successfully established in the soil after hardening. The survival percentage of rooted plants on soil was found to be 85%. This result will facilitate the conservation and propagation of the important medicinal herb Physalis angulata L.

scholarly journals Induksi Mutasi Kalus Embriogenik Gandum (Triticum aestivum L.) melalui Iradiasi Sinar Gamma untuk Toleransi Suhu Tinggi

2015 ◽  
Vol 43 (1) ◽  
pp. 36 ◽  
Author(s):  
Ryan Budi Setiawan ◽  
Nurul Khumaida ◽  
Diny Dinarti
Keyword(s):  
High Temperature ◽  
Embryogenic Callus ◽  
Genetic Improvement ◽  
In Vitro Selection ◽  
Gamma Ray ◽  
Triticum Aestivum L ◽  
Temperature Tolerance ◽  
Randomized Design ◽  
Gamma Ray Irradiation

Mutation techniques through gamma ray irradiation is useful to support breeding programs for genetic improvement of wheat. Genetic improvement on tolerance to high temperatures is necessary for development of wheat in Indonesia. The purpose of this study was to determine the level of radiosensitivity to be used as the basis for the induction of mutations by gamma ray irradiation on embryogenic callus to obtain putative mutants with high temperature tolerance. Explants used were embryogenic callus cultured on MS medium containing 30 g L-1 sukrosa, 2 g L-1 gelrite, 2 mg L-1 2.4D and 1 mg L-1 picloram. Culture incubated for 6 weeks with temperature 20±4 oC in the room culture. Five irradiation doses (10, 20, 30, 40 and 50 Gy) were used in radiosensitivity testing. A factorial, completely randomized design was applied to the experiment. The first factor was selection temperature with three levels (25, 30 and 35 oC), and the second factor was doses of gamma ray iradiation with three levels (10, 20, and 30 Gy). The result showed that radiosensitivity levels varied among varieties, LD20: 7.79 to 18.96 Gy and LD50: 24.29-33.63 Gy. Selayar variety which has highest sensitivity value compared with Dewata and Nias. Increasing doses of iradiation and temperature decrease survival of embryogenic callus, number of embryos, and percentage of germinated plantlets. Based on in vitro selection using high temperature (25, 30, and 35 oC), the obtained 19 putative mutants were derived from embryos that appear on the surface of embryogenic callus survival after irradiation and high temperature selection.<br />Keywords: in vitro selection, putative mutant, radiosensitivity, somatic embryo

scholarly journals In vitro Development of Cauliflower Synthetic Seeds and Development of Plantlets In vivo

2014 ◽  
Vol 24 (1) ◽  
pp. 27-36 ◽  
Author(s):  
Zahida Qamar ◽  
Md. Belal Hossain ◽  
Idrees A. Nasir ◽  
Bushra Tabassum ◽  
Tayyab Husnain
Keyword(s):  
Cell Suspension ◽  
Somatic Embryos ◽  
Cell Suspension Cultures ◽  
Synthetic Seeds ◽  
Embryogenic Calli ◽  
Globular Cells ◽  
Vitro Development ◽  
Direct Use

Synthetic seeds of cauliflower cv. Chillout were developed by encapsulating mature somatic embryos in neutral gel media. Somatic embryos were obtained by optimizing callus and cell suspension cultures of cauliflower. Friable, yellowish embryogenic calli were obtained on MS supplemented with 2 mg/l  2,4-D and 0.5 mg/l BAP using hypocotyl as explants, while calli were regenerated in media consisting of 5 mg/l BAP, 2 mg/l Kn and 6 mg/l GA3. Somatic embryo-genesis was induced in cell suspension culture where auxins were removed in successive steps triggering  conversion of globular cells into the heart, torpedo stage (71%) and finally into cotyledonary/somatic embryos (28%). The mature somatic embryos were encapsulated by mixing mature cell suspension with sodium alginate and calcium chloride mixture (1 : 4). Developed synthetic seeds germinated into complete plantlets when placed in neutral gel media.  Germination efficiency of synthetic seeds decreased to about 50 per cent after 12 weeks of storage at 4ºC followed by a rapid decrease to zero per cent after 16 weeks. It was also observed that cauliflower plantlets from synthetic seeds survived successfully when transferred to soil demonstrating  that cauliflower synthetic seeds is a promising step towards their  in vivo direct use. Plant Tissue Cult. & Biotech. 24(1): 27-36, 2014 (June) D. O. I. http://dx.doi.org/10.3329/ptcb.v24i1.19193

scholarly journals Efficient Somatic Embryogenesis and Plant Regeneration from Immature Embryos of Tapiscia sinensis Oliv., an Endemic and Endangered Species in China

HortScience ◽  
2014 ◽  
Vol 49 (12) ◽  
pp. 1558-1562 ◽  
Author(s):  
Yuyu Wang ◽  
Faju Chen ◽  
Yubing Wang ◽  
Xiaoling Li ◽  
Hongwei Liang
Keyword(s):  
Somatic Embryogenesis ◽  
Plant Regeneration ◽  
Activated Charcoal ◽  
Somatic Embryos ◽  
Basal Medium ◽  
Immature Embryos ◽  
Zygotic Embryos ◽  
Naphthaleneacetic Acid ◽  
Embryogenic Calli ◽  
Induction Rate

High-frequency somatic embryogenesis and plant regeneration were achieved from immature cotyledonary-stage embryos in the endangered plant, Tapiscia sinensis Oliv. Plant growth regulators with different concentrations and combinations on embryogenesis capacity were studied. The optimal explants for in vitro somatic embryogenesis were immature embryos in T. sinensis. A high callus induction rate of 100% was achieved on Murashige and Skoog (MS) basal medium supplemented with 1.0 mg·Ll−1 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.5% (w/v) activated charcoal. Alternatively, a high induction rate (96.16%) of somatic embryogenesis was obtained on MS basal medium supplemented with the combination of 0.05 mg·L−1 α-naphthaleneacetic acid (NAA) and 0.2 mg·L−1 6-benzylaminopurine (6-BA), and somatic embryos proliferated fastest on the mentioned medium supplemented with 0.5% (w/v) activated charcoal and 3% (w/v) sucrose, inoculation of explants proliferating 21 times in the 23-day subculture. Of the 100 plantlets transferred to field after the acclimation, 95 (95%) survived. Based on the histocytological observations, the development of somatic embryos was similar to that of zygotic embryos. There were two accumulation peaks of starch grains in the embryogenic calli and in the globular-stage embryos, both closely related to the energy supply, and the embryoids were of multicelluar origin.

scholarly journals Somatic Embryogenesis in Centella asiatica, a Valuable Medicinal Plant

HortScience ◽  
2004 ◽  
Vol 39 (4) ◽  
pp. 859C-859
Author(s):  
Nirmal Joshee* ◽  
Bipul K. Biswas ◽  
Anand K. Yadav
Keyword(s):  
Somatic Embryos ◽  
Bacterial Contamination ◽  
Developmental Stages ◽  
Basal Medium ◽  
Centella Asiatica ◽  
Perennial Herb ◽  
Culture Techniques ◽  
Nodular Callus ◽  
Further Development

Centella asiatica L. (Apiaceae family), also called `Indian Pennywort,' is a prostrate, faintly aromatic, and stoloniferous perennial herb with long petiolated leaves. In the Ayurvedic medicine, it is reputed as a nervine tonic along with antibacterial, antifeedant, antileprotic and wound healing properties. Centella contains glycosides, indocentelloside, brahmoside, and asiaticoside. Its leaves are rich in carotenoids and vitamins B and C. In vitro culture techniques which offer a viable tool for mass propagation of plants have recently become increasingly popular for conservation of rare, endangered and threatened medicinal plants germplasm. Centella tissue culture has been reported to experience high incidences of microbial contamination which drastically reduces survival of explants. Thus, the main purpose of this study was to develop an efficient micropropagation technique for Centella asiatica to reduce explant contamination and rapidly disseminate superior clones for research and production. Here we present induction and further development of somatic embryos, using Centella stolons as explants. Somatic embryos were induced in response to 2,4-D shock on MS medium. Initially, somatic embryos appeared as highly nodular callus and eventually developed into somatic embryos that exhibited globular, heart shaped and cotyledonary stages. After auxin shock, cultures were regularly transferred to MS basal medium where somatic embryos completed various developmental stages and then germinated to give rise to new plantlets. In this presentation, we will demonstrate complete protocols for the successful sterilization of Centella explants prepared from plants that had abundance of fungal and bacterial contamination.

scholarly journals PENGARUH SUHU INKUBASI TERHADAP PERTUMBUHAN DAN PERKEMBANGAN EMBRIO SOMATIK PURWOCENG (Pimpinella pruatjan Molk.)

2020 ◽  
Vol 16 (2) ◽  
pp. 56
Author(s):  
NUR AJIJAH ◽  
IRENG DARWATI ◽  
YUDIWANTI YUDIWANTI ◽  
ROOSTIKA ROOSTIKA
Keyword(s):  
Somatic Embryos ◽  
Growth And Development ◽  
In Vitro Selection ◽  
Incubation Temperature ◽  
Effect Of Temperature ◽  
Fresh Weight ◽  
Temperature Growth ◽  
Weight Increment ◽  
Shoot Number

<p>ABSTRAK</p><p>Purwoceng (Pimpinella pruatjan Molk. atau P. alpina KDS.) merupakan salah satu tanaman obat asli Indonesia endemik dataran tinggidan pada saat ini dibudidayakan secara terbatas di Dataran Tinggi Dieng.Salah satu upaya yang dapat ditempuh untuk memperluas arealpengembangan tanaman ini adalah melalui perakitan varietas tolerandataran rendah atau menengah, yang antara lain dapat diperoleh melaluipendekatan seleksi ketahanan terhadap suhu tinggi yang dapat dilakukansecara in vitro. Pengaruh cekaman suhu tinggi terhadap pertumbuhan danperkembangan purwoceng secara in vitro sejauh ini belum diketahui.Penelitian bertujuan untuk mengetahui pengaruh suhu inkubasi terhadappertumbuhan dan perkembangan embrio somatik purwoceng secara invitro. Penelitian dilaksanakan di laboratorium kultur jaringan BalaiPenelitian Tanaman Obat dan Aromatik (Balittro) Bogor mulai Oktober2007 – Maret 2008. Embrio somatik purwoceng diinduksi dari eksplandaun aseptik. Embrio somatik fase globuler yang terbentuk dipergunakansebagai eksplan kemudian diinkubasi pada tiga taraf suhu ruang yaitu 17,3± 0,5ºC (kontrol), 23,3 ± 2,1ºC, dan 32,8 ± 1,7ºC selama 3 bulan dengansub kultur setiap bulan sampai terbentuk planlet/tunas. Pengamatandilakukan terhadap peubah pertumbuhan dan perkembangan eksplanembrio somatik yang meliputi penambahan bobot segar eksplan,persentase eksplan yang membentuk tunas, jumlah tunas yang terbentukper eksplan serta persentase eksplan hidup. Hasil penelitian menunjukkanbahwa suhu inkubasi berpengaruh nyata terhadap semua peubah yangdiamati. Rata-rata  penambahan  bobot  segar,  persentase  eksplanmembentuk tunas, jumlah tunas per eksplan dan persentase eksplan hidupsemakin menurun dengan semakin meningkatnya suhu inkubasi. Suhu 32,8± 1,7ºC memberikan pengaruh penghambatan yang nyata terhadappertumbuhan dan perkembangan embrio somatik purwoceng dibandingkansuhu kontrol dan 23,3 ± 2,1ºC sejak periode inkubasi 1 bulan. Sedangkansuhu 23,3 ± 2,1ºC baru memberikan pengaruh penghambatan yang nyatasetelah periode inkubasi 3 bulan.</p><p>Kata kunci : Pimpinella pruatjan, embrio somatik, suhu, pertumbuhan</p><p>ABSTRACT</p><p>Effect of temperature incubation on growth and de-velopment of Purwoceng (Pimpinella pruatjan Molk.)somatic embryos</p><p>Purwoceng (Pimpinella pruatjan Molk., P. alpina KDS.) is one ofIndonesian medicinal plants. It is high altitude endemic species which iscurrently cultivated on a limited areas in the Dieng Plateau. One effort toexpand purwoceng cultivation areas is through the assembly ofpurwoceng varieties tolerant to low or medium altitude, among others, canbe obtained through the approach of selection for high temperaturetolerance that can be done by in vitro selection. How high temperaturestress influencing the growth and development of purwoceng somaticembryos is not known yet. The research aimed at determining theinfluence of incubation temperature on the growth and development ofpurwoceng somatic embryos. The research was conducted at tissue culturelaboratory of Indonesian Medicinal and Aromatic Crops Research Institute(IMACRI) from October 2007 – March 2008. Purwoceng somaticembryos induced from aseptic leaves incubated at three levels of roomtemperature i.e. 17.3 ± 0.5 º C (control), 23.3 ± 2.1 ºC, and 32.8 ± 1.7 º Cfor 3 months with a subculture every month. Variables observed wereexplants fresh weight increment, percentage of explants forming shoot,number of shoot per explants, and percentage of survive explants. Theresult showed that the average of explants fresh weight increment,percentage of explants forming shoot, number of shoot per explants, andpercentage of survive explants decreased with the increase of temperature.The growth and development of purwoceng somatic embryos weresignificantly inhibited at the temperature of 32.8 ± 1.7ºC since one monthafter incubation, while the inhibition of temperature of 23.3 ± 2.1 ºC wasnot significant except after 3 month of incubation.</p><p>Key words: Pimpinella pruatjan, somatic embryos, temperature, growth</p>

scholarly journals Slow-growth conservation- cryopreservation and Analysis of genetic stability of in vitro regenerated Coelogyne Nervosa A.Rich A endemic orchid

2021 ◽  
Vol 12 (2) ◽  
pp. 1695-1698
Author(s):  
Chithra Devi B.S. ◽  
Sangilimuthu A ◽  
Daanya V
Keyword(s):  
Somatic Embryos ◽  
Tamil Nadu ◽  
Slow Growth ◽  
Basal Medium ◽  
Genetic Changes ◽  
Storage Media ◽  
Inter Simple Sequence Repeats ◽  
Growth Preservation ◽  
Simple Sequence

Coelogyne Nervosa A. Rich., an endangered medicinal epiphyte (lithophyte) is a strict endemic of the Western Ghats distributed across Karnataka, Kerala and Tamil Nadu. KC medium supplemented with 2,4-D is best suited for direct embryo induction from 80 days old protocorms. Hormone-free KC basal medium supported the conversion of somatic embryos into complete plantlets. For the medium-term storage experiment, somatic embryos were placed on storage media containing various growth inhibitors (such as different levels of chlormequat, maleic hydrazide and paclobutrazol). They were stored for ten months to assess the percentage of survival. The surviving shoots were transferred to the recovery medium for eight weeks. At 8°C, the highest survival rate was observed with ABA at 0.2 mg L−1. For cryopreservation: droplet-vitrification best results were obtained when the somatic embryos, after 20 days of subculture were immersed in an osmoprotectant solution. Osmo-protected samples were immersed in PVS2 (15% ethylene glycol and 15% DMSO) at 0°C for 30 min. Pre-prepared Somatic embryos were plunged into cryovials for one day. Regrowth levels were as high as 70%. DNA samples extracted from leaves of the mother plant, somatic embryos without storage, regenerates of slow growth preservation and cryopreservation. The samples were analysed by DNA based molecular marker – inter simple sequence repeats (ISSR) and found no genetic changes during the cryopreservation process.

Sign in / Sign up

Export Citation Format

Share Document