scholarly journals In Vitro Selection of a Salt-tolerant Embryogenic Line in Habanero Pepper (Capsicum chinense Jacq.)

HortScience ◽  
2011 ◽  
Vol 46 (12) ◽  
pp. 1666-1671 ◽  
Author(s):  
Daniela Solís-Marroquín ◽  
Carlos Alberto Lecona-Guzmán ◽  
Adriana Canto-Flick ◽  
Jericó Jabín Bello-Bello ◽  
Nancy Santana-Buzzy ◽  
...  
Keyword(s):  
Salt Stress ◽  
Somatic Embryos ◽  
Growth And Development ◽  
In Vitro Selection ◽  
Lethal Dose ◽  
Intermediate Stage ◽  
Cellular Mechanisms ◽  
Stage Of Development ◽  
Habanero Pepper

The effect of NaCI salinity on growth and development of somatic embryos of Habanero pepper was examined. Addition of 75 and 100 mm NaCI into the medium greatly increased the growth and development of somatic embryos and both of these concentrations favored the proliferation of somatic embryos. However, supplementation of 200 and 300 mm NaCI to the medium showed a negative effect on the growth and development of somatic embryos. Concentration increases of NaCl provoked a significant reduction of the embryos survival rate with the average lethal dose (46%) being registered in the treatment of 100 mm. Furthermore, a lower tolerance to salt stress (NaCl) was observed in deformed somatic embryos. Concentrations of 200 and 300 mm NaCl significantly delayed development in the surviving embryos in both treatments. These embryos remained at the globular stage throughout culture time. At 75 mm NaCl, most of the embryos were observed in the torpedo stage. However, the embryos exposed to 100 mm NaCl were observed mainly in globular and cotiledonar stages. It is quite likely that the transition from one intermediate stage of development to another occurs rapidly. With the exception of the concentration at 300 mm NaCl, salt stress stimulated embryonic germination, particularly at 100 mm NaCl. The content of proline in somatic embryos increased substantially in response to salinization. The results suggest that somatic embryos of C. chinense can tolerate concentrations of NaCl up to 100 mm without their development being affected. Moreover, they have sufficient cellular mechanisms to tolerate salinity at relatively higher levels.

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 Characterization of Plant Growth-Promoting Traits and Inoculation Effects on Triticum durum of Actinomycetes Isolates under Salt Stress Conditions

Soil Systems ◽  
2021 ◽  
Vol 5 (2) ◽  
pp. 26
Author(s):  
Rihab Djebaili ◽  
Marika Pellegrini ◽  
Massimiliano Rossi ◽  
Cinzia Forni ◽  
Maria Smati ◽  
...  
Keyword(s):  
Acetic Acid ◽  
Salt Stress ◽  
Plant Growth ◽  
Durum Wheat ◽  
Growth And Development ◽  
Indole Acetic Acid ◽  
Greenhouse Experiment ◽  
Hydrocyanic Acid ◽  
In Vitro Assays

This study aimed to characterize the halotolerant capability, in vitro, of selected actinomycetes strains and to evaluate their competence in promoting halo stress tolerance in durum wheat in a greenhouse experiment. Fourteen isolates were tested for phosphate solubilization, indole acetic acid, hydrocyanic acid, and ammonia production under different salt concentrations (i.e., 0, 0.25, 0.5, 0.75, 1, 1.25, and 1.5 M NaCl). The presence of 1-aminocyclopropane-1-carboxylate deaminase activity was also investigated. Salinity tolerance was evaluated in durum wheat through plant growth and development parameters: shoot and root length, dry and ash-free dry weight, and the total chlorophyll content, as well as proline accumulation. In vitro assays have shown that the strains can solubilize inorganic phosphate and produce indole acetic acid, hydrocyanic acid, and ammonia under different salt concentrations. Most of the strains (86%) had 1-aminocyclopropane-1-carboxylate deaminase activity, with significant amounts of α-ketobutyric acid. In the greenhouse experiment, inoculation with actinomycetes strains improved the morpho-biochemical parameters of durum wheat plants, which also recorded significantly higher content of chlorophylls and proline than those uninoculated, both under normal and stressed conditions. Our results suggest that inoculation of halotolerant actinomycetes can mitigate the negative effects of salt stress and allow normal growth and development of durum wheat plants.

Nodulation under salt stress of alfalfa lines obtained after in vitro selection for osmotic tolerance

Plant Science ◽  
2003 ◽  
Vol 165 (4) ◽  
pp. 887-894 ◽  
Author(s):  
Dimitar Djilianov ◽  
Els Prinsen ◽  
Sevgi Oden ◽  
Henry van Onckelen ◽  
Joachim Müller
Keyword(s):  
Salt Stress ◽  
In Vitro Selection ◽  
Osmotic Tolerance ◽  
Selection For

scholarly journals Efficient Transformation of Somatic Embryos and Regeneration of Cork Oak Plantlets with a Gene (CsTL1) Encoding a Chestnut Thaumatin-Like Protein

2021 ◽  
Vol 22 (4) ◽  
pp. 1757
Author(s):  
Vanesa Cano ◽  
Mᵃ Teresa Martínez ◽  
José Luis Couselo ◽  
Elena Varas ◽  
Francisco Javier Vieitez ◽  
...  
Keyword(s):  
Somatic Embryos ◽  
Transformation Efficiency ◽  
Phytophthora Cinnamomi ◽  
Lethal Dose ◽  
Cork Oak ◽  
Pcr Analysis ◽  
Qpcr Analysis ◽  
Bacteriostatic Agent ◽  
Transgenic Lines

We present a reproducible procedure for transforming somatic embryos of cork oak with the CsTL1 gene that codes for a thaumatin-like protein, in order to confer tolerance to Phytophthora cinnamomi. Different concentrations/combinations of the antibiotics carbenicillin and cefotaxime, as bacteriostatic agents, and kanamycin, as a selective agent, were tested. A lethal dose of 125 mg/L kanamycin was employed to select transgenic somatic embryos, and carbenicillin was used as a bacteriostatic agent at a concentration of 300 mg/L, which does not inhibit somatic embryo proliferation. The transformation efficiency was clearly genotype-dependent and was higher for the TGR3 genotype (17%) than for ALM80 (4.5%) and ALM6 (2%). Insertion of the transgenes in genomic DNA was confirmed by PCR analysis, whereas expression of the CsTL1 gene was evaluated by semi-quantitative real-time PCR (qPCR) analysis. A vitrification treatment successfully cryopreserved the transgenic lines generated. The antifungal activity of the thaumatin-like protein expressed by the gene CsTL1 was evaluated in an in vitro bioassay with the oomycete P. cinnamomi. Of the eight transgenic lines analyzed, seven survived for between one or two times longer than non-transgenic plantlets. Expression of the CsTL1 gene and plantlet survival days were correlated, and survival was generally greater in plantlets that strongly expressed the CsTL1 gene.

scholarly journals In vitro selection of winter triticale for salt resistance

1970 ◽  
pp. 247-251
Author(s):  
S. V. Pykalo ◽  
O. V. Dubrovna ◽  
O. A. Demydov
Keyword(s):  
Salt Stress ◽  
Cell Lines ◽  
In Vitro Selection ◽  
High Survival ◽  
Winter Triticale ◽  
Step Type ◽  
Callus Lines ◽  
Selection Of

Aim. To obtain of cell lines and plant-regenerants of winter triticale resistant for salt stress the in vitro selection was carried out. Methods. In order to select resistant to salt stress forms of triticale the efficiency of using direct and step-type in vitro selection with application of selective system based on sodium chloride has been investigated. Results. The direct and step-type in vitro selection was conducted and the selection of callus lines of triticale being resistant to simu-lated salinity was carried out. As a result, from line 38/1296 and variety Obriy respectively, 5 and 4 resistant callus lines were identified that had a high survival rate on the selective medium with 1.2 % NaCl and maintained morphogenetic potential. From the resistant lines plant regenerants were induced and their rearing, rooting and transfer to in vivo condi-tions were optimized. Conclusions. A step-type in vitro selection was more effective, because resulted from the selec-tion more resistant callus forms were identified. First cell lines of winter triticale with resistance to salt stress were de-rived.Keywords: Triticale, in vitro selection, callus, salt stress, resistance.

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 Obtaining Salt Stress-Tolerant Eggplant Somaclonal Variants from In Vitro Selection

Plants ◽  
2021 ◽  
Vol 10 (11) ◽  
pp. 2539
Author(s):  
Sami Hannachi ◽  
Stefaan Werbrouck ◽  
Insaf Bahrini ◽  
Abdelmuhsin Abdelgadir ◽  
Hira Affan Siddiqui ◽  
...  
Keyword(s):  
Ascorbic Acid ◽  
Salt Stress ◽  
Water Content ◽  
Acid Content ◽  
In Vitro Selection ◽  
Callus Tissue ◽  
Salt Content ◽  
Ascorbic Acid Content ◽  
Control Line

An efficient regeneration protocol was applied to regenerate shoots on salt stress-tolerant calli lines of aubergine (Solanum melongena). These NaCl-tolerant cell lines were obtained by two different methods. On the one hand, the developed callus tissue was transferred to a medium with a continuous salt content of 40, 80, 120, or 160 mM NaCl. On the other hand, the callus tissue was subjected to a stepwise increasing salinity to 160 mM NaCl every 30 days. With the second method, calli which could be selected were characterized by compact growth, a greenish color, and absence of necrotic zones. When grown on salt-free medium again, NaCl-tolerant calli showed a decline in relative growth rate and water content in comparison to the control line. This was more obvious in the 120 mM NaCl-tolerant callus. Lipid peroxidase activity increased in 40 and 80 mM NaCl-tolerant calli; yet did not increase further in 120 mM-tolerant callus. An increase in ascorbic acid content was observed in 80 and 120 mM NaCl-tolerant calli compared to the 40 mM NaCl-tolerant lines, in which ascorbic acid content was twice that of the control. All NaCl-tolerant lines showed significantly higher superoxide dismutase (SOD) (208–305–370 µmol min−1 mg−1 FW) and catalase (CAT) (136–211–238 µmol min−1 mg−1 FW) activities compared to control plants (231 and 126 µmol min−1 mg−1 FW). Plants were regenerated on the calli lines that could tolerate up to 120 mM NaCl. From the 32 plants tested in vitro, ten plants with a higher number of leaves and root length could be selected for further evaluation in the field. Their high salt tolerance was evident by their more elevated fresh and dry weight, their more increased relative water content, and a higher number and weight of fruits compared to the wild-type parental control. The presented work shows that somaclonal variation can be efficiently used to develop salt-tolerant mutants.

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