Mutagenesis of in vitro explants of Coffea spp. to induce fungal resistance.

2021 ◽  
pp. 344-352
Author(s):  
Souleymane Bado ◽  
Fatemeh Maghuly ◽  
Vitor Varzea ◽  
Margit Laimer

Abstract Coffee is one of the most valuable commodity tree crops worldwide. However, it suffers from several devastating diseases and pests, for example coffee leaf rust and coffee berry borer, whose impact is being amplified by changing climatic conditions. Development of new adapted varieties remains a laborious effort by conventional breeding due to the long juvenile period in tree crops. Plant cell/tissue culture represents the ultimate method to produce large amounts of true-to-type healthy plants and of explants for mutation breeding. In fact, mutation induction combined with in vitro cell/tissue culture techniques has proved to be effective for developing improved cultivars of perennial crops. Prior to mutation breeding, cell and tissue radiosensitivity tests to various mutagens need to be performed, so that optimal treatments can be applied for large population development. Thus, different in vitro explants (plantlet, leaf, callus, embryogenic callus, globular and torpedo stage embryos) of Coffea arabica and Coffea canephora were exposed to different gamma-ray doses (0, 10, 15, 20, 40, 60 and 80 Gy). After 9-21 weeks incubation, a radiosensitivity test was conducted on the different explants and LD50 doses corresponding to 50% of viability or survival of callus, embryogenic callus, globular and torpedo stage embryos and 50% growth reduction (GR50) of shoot were also determined. Callus explants showed a relatively high radio-resistance (LD30-LD50 50-100 Gy) in comparison with entire plantlets or embryos (LD30-GR50 8-46 Gy). Globular embryo development into plantlets and also leaf area of irradiated plantlets were more severely affected by irradiation than other explants. It was possible to confirm the relative radio-resistance of unicellular explants compared with multicellular explants. Estimation of optimal mutation induction dosage range for various in vitro explants is important for tree crops, especially for coffee improvement.

2020 ◽  
Vol 58 (4) ◽  
pp. 393
Author(s):  
Hoàng Thanh Tùng ◽  
Luan Quoc Vu ◽  
Nhut Duong

Paphiopedilum orchids are one of the most popular and rare orchid genera sold and exhibited as pot plants and cut flowers. Their wild populations are under the threat of extinction as a result of over-collection and loss of suitable habitats. Reduction in their commercial value through large-scale propagation in vitro is a preferable option to reduce pressure from illegal collection, to attempt at meeting commercial needs and to re-establish these threatened orchid species back into the wild. Although they are commercially propagated via seed germination in vitro, Paphiopedilum are considered to be difficult to propagate in vitro, especially by plant regeneration from tissue culture. This paper aims to provide the most important techniques on Paphiopedilum propagation mainly including plant, cell, tissue and organ culture techniques applied to in vitro propagation of Paphiopedilum and to emphasize the importance of further improving tissue culture protocols from ex vitro-derived explants of mature plants.


2016 ◽  
Vol 8 (1) ◽  
pp. 38 ◽  
Author(s):  
Endang G. Lestari

<p>Mutation-based<br />plant improvement, which changes one or a few specific<br />traits of a cultivar, can contribute to crop improvement.<br />Tissue culture increases the efficiency of mutagenic<br />treatment to induce variations. In vitro culture in<br />combination with induced mutation can speed up the<br />breeding program by generating variability, followed by<br />selection and multiplication of the desired genotypes. In<br />many vegetative propagated crops, mutation induction in<br />combination with in vitro culture techniques can be the<br />most effective method for plant improvement. In seed<br />propagated species, the application of mutation coupled<br />with doubled haploid systems seems to be highly promising<br />in crop improvement. This approach speeds up the breeding<br />program through generation of variability followed by<br />selection of homozygousity and rapid multiplication of<br />desired genotypes.</p>


2015 ◽  
Vol 77 (24) ◽  
Author(s):  
Siti Suhaila A. Rahman ◽  
Norwati Muhammad ◽  
Nor Hasnida Hassan ◽  
Haliza Ismail ◽  
Nazirah Abdullah ◽  
...  

Neolamarckia cadamba (kelempayan) is a multipurpose and fast growing timber species. The tree is grown for timber, paper-making and as ornamental plant. It is reported that its barks and leaves possesed medicinal values and its flowers are used in perfumes. The species is also known to be suitable for plywood, packing case, toys and short-fibred pulp. Therefore, mass production of high quality planting material of N. cadamba is important to support plantation program of this species. Here we presented mass production of N. cadamba through tissue culture techniques. Nodal segments derived from in vitro germinated seeds were used and induced direct organogenesis to produce shoots and roots using MS media (1962) and plant growth regulators (BAP and IBA) that are relatively cheaper than previously used methods. The tissue culture technique of N. cadamba developed may help in ensuring supply of planting materials that are feasible for commercial plantation purposes.


2002 ◽  
Vol 68 (2) ◽  
pp. 656-660 ◽  
Author(s):  
Stephen L. Dobson ◽  
Eric J. Marsland ◽  
Zoe Veneti ◽  
Kostas Bourtzis ◽  
Scott L. O'Neill

ABSTRACT Maternally transmitted bacteria of the genus Wolbachia are obligate, intracellular symbionts that are frequently found in insects and cause a diverse array of reproductive manipulations, including cytoplasmic incompatibility, male killing, parthenogenesis, and feminization. Despite the existence of a broad range of scientific interest, many aspects of Wolbachia research have been limited to laboratories with insect-rearing facilities. The inability to culture these bacteria outside of the invertebrate host has also led to the existing bias of Wolbachia research toward infections that occur in host insects that are easily reared. Here, we demonstrate that Wolbachia infections can be simply established, stably maintained, and cryogenically stored in vitro using standard tissue culture techniques. We have examined Wolbachia host range by introducing different Wolbachia types into a single tissue culture. The results show that an Aedes albopictus (Diptera: Culicidae) cell line can support five different Wolbachia infection types derived from Drosophila simulans (Diptera: Drosophilidae), Culex pipiens (Culicidae), and Cadra cautella (Lepidoptera: Phycitidae). These bacterial types include infection types that have been assigned to two of the major Wolbachia clades. As an additional examination of Wolbachia host cell range, we demonstrated that a Wolbachia strain from D. simulans could be established in host insect cell lines derived from A. albopictus, Spodoptera frugiperda (Lepidoptera: Noctuidae), and Drosophila melanogaster. These results will facilitate the development of a Wolbachia stock center, permitting novel approaches for the study of Wolbachia infections and encouraging Wolbachia research in additional laboratories.


2021 ◽  
Vol 11 (2) ◽  
pp. 114
Author(s):  
Nurkapita Nurkapita ◽  
Riza Linda ◽  
Zulfa Zakiah

(Article History: Received February 18, 2021; Revised April 27, 2021; Accepted May 19, 2021) ABSTRAKPerkembangbiakan anggrek secara generatif alami membutuhkan bantuan jamur mikoriza untuk perkecambahan biji, sedangkan usaha perbanyakan konvensional memerlukan waktu lama untuk memperoleh tanaman dalam jumlah banyak. Salah satu alternatif untuk perbanyakan anggrek hitam (Coelogyne pandurata Lindl.) adalah melalui multiplikasi tunas anggrek secara in vitro. Tujuan penelitian adalah untuk membuktikan pengaruh pemberian NAA (Naphthalene Acetic Acid) dan ekstrak biji jagung (Zea mays) terhadap multiplikasi tunas anggrek hitam. Penelitian dilakukan di Laboratorium Kultur Jaringan Jurusan Biologi Fakultas Matematika dan Ilmu Pengetahuan Alam Universitas Tanjungpura Pontianak. Penelitian ini menggunakan Rancangan Acak Lengkap (RAL) pola faktorial dengan dua faktor perlakuan. Faktor pertama adalah NAA terdiri dari 5 taraf konsentrasi yaitu A0 (0 M/ kontrol) A1 (10-7 M), A2 (10-6 M), A3 (5x10-7 M) dan A4 (5x10-6 M ) dan faktor ekstrak biji jagung (B) dengan 5 taraf konsentrasi yaitu B0 (0%), B1 (2,5%), B2 (5%); B3 (7,5%) dan B4 (10%). Pemberian kombinasi NAA dan ekstrak biji jagung berpengaruh nyata terhadap semua parameter yaitu jumlah tunas, jumlah daun, dan tinggi tunas. Hasil terbaik rerata jumlah tunas pada perlakuan A4+B4 yaitu 5x10-6M NAA+10% ekstrak biji jagung. Hasil terbaik pada rerata jumlah daun pada perlakuan A2+B2 yaitu 5x10-7M NAA+5% ekstrak biji jagung dan hasil terbaik pada rerata tinggi tunas pada perlakuan A1+B1 yaitu 10-7M NAA+2,5% ekstrak biji jagung.Kata Kunci: multiplikasi; tunas anggrek hitam; ekstrak biji jagung; NAA. ABSTRACTGenerative reproduction of orchid plants it takes a requires the help of mycorriza mushrooms for seed germination, whereas conventional propagation business takes a long time to obtain large quantities of plants. One alternative to the propagation black orchids (Coelogyne pandurata Lindl.) is required through tissue culture techniques. The purpose of this study is to find the influence and concentration corn seed extract (Zea mays) and NAA (Naphthalene Acetic Acid) on the multiplication black orchids. This research was conducted in the tissue culture laboratory Biology Department Faculty of Mathematics and Natural Sciences Tanjungpura University Pontianak. The study used a Complete Randomized Design (RAL) of factorial patterns with two treatment factors. The first factor is that the NAA consists of 5 concentration levels  A0 (0 M) A1 (10-7 M), A2 (10-6 M), A3 (5x10-7 M) and A4 (5x10-6 M ) and the second factor is that corn seed extract of 5 levels concentratio B0(0%), B1 (2,5%), B2 (5%); B3 (7,5%) and B4 (10%). The administration NAA and corn seed extract in combination has a real effect on all parameters namely the number shoots, the number leaves, and the height shoots. The best results where the average number of shoots in the treatment of A2+B2 namely 5x10-6M NAA + 10% corn seed extract. The best results average number of leaves in the treatment  A2+B2 namely 5x10-7M NAA + 5% corn seed extract and in the best results for shoot height in the treatment of A1+B1 namely 10-7M NAA + 2.5% corn seed extract.Keywords: Multiplication; black orchid’s shoot; corn  seed extract; NAA


Agrin ◽  
2019 ◽  
Vol 22 (1) ◽  
pp. 10
Author(s):  
Asih K. Karjadi ◽  
Neni Gunaeni

Tanaman bawang merah (Allium ascolonicum L) termasuk dalam genus Allium sp yang diperbanyak secaravegetatif melalui umbi. Perbanyakan benih bawang sudah dilakukan secara in vitro (konvensional), untuk tujuanpeningkatan mutu atau hanya perbanyakan tanaman . Penelitian dilaksanakan di Laboratorium Kultur JaringanBalai Penelitian Tanaman Sayuran, pada bulan Februari sampai Agustus 2014. Untuk menghasilkan tanamanbebas penyakit terutama virus dapat digunakan teknik kultur jaringan yang dikombinasikan dengan perlakuanpemanasan. Penelitian bertujuan untuk melihat pengaruh pemanasan dan sumber eksplan terhadap pertumbuhandan perkembangan eksplan bawang merah.. Perlakuan pemanasan bahan eksplan bawang merah dilakukan secarabertahap selama 4 minggu, masing-masing 1 minggu untuk suhu (30, 33, 35 dan 37 oC). Media yang dipergunakanuntuk penumbuhan eksplan adalah MS + MS vits + sucrose 30 g/l + IAA 2 mg/l + Kinetin 2 mg/l + GA3 0.01mg/l agar gelgro 2 g/l pH 5.7. Penelitian terdiri dari 2 kegiatan yaitu Perlakuan pada cv. Pikatan, dan pada cv.Bima Brebes. Sebagai eksplan dipergunakan yaitu (1/3 bulb/ umbi) dan (shoot tip/ jaringan meristematik denganbeberapa daun primordia). Perlakuan eksplan yang digunakan yaitu tanpa pemanasan dan dengan pemanasan.Pertumbuhan dan perkembangan dari planlet diamati pada penelitian ini. Hasil dari penelitian menunjukkanbahwa perlakuan pemanasan bahan eksplan bawang merah cv. Bima Brebes, belum menurunkan persentase planletyang terinfeksi virus. Eksplan (1/3 bulb) dan (shoot tip), mempunyai pertumbuhan eksplan diatas 50%. Umumnyasemakin kecil eksplan persentase planlet abnormal semakin tinggi. Kontaminasi kultur umumnya disebabkanbakteri dan jamur yang terbawa dari eksplan (endogen). Perlakuan pemanasan bahan eksplan bawang merah secaravisual tidak berpengaruh pada persentase pertumbuhan dan persentase kultur terkontaminasi.Kata kunci: Bawang merah, Allium ascolonicum L, pemanasan, asal eksplanABSTRACTRed onion plants (Allium ascolonicum L) are included in the genus Allium sp which is propagatedvegetatively through tubers. Propagation of onion seeds has been done in vitro (conventionally), for the purposeof quality improvement or only plant propagation. The research was carried out at the Tissue Culture Laboratory,Balai Penelitian Tanaman Sayuran (Balitsa) from February to August 2014. To produce plants free of mainly viraldiseases can be used tissue culture techniques combined with heating treatment. Research aims were to look atthe effect of heating and explants on the growth and development of red onion explants. The treatment of the onionexplants was heated gradually for 4 weeks, 1 week each for temperatures (30, 33, 35 and 37 oC). The media usedfor the growth of explants is MS + MS vits + sucrose 30 g / l + IAA 2 mg / l + Kinetin 2 mg / l + GA3 0.01 mg / lso that gelgro 2 g / l pH 5.7. The study consisted of 2 activities, namely, treatment at cv. Pikatan, and at cv. BimaBrebes. As explants it is used (1/3 bulb / tuber) and (shoot tip / meristematic tissue with several leaves ofprimordia). The explants treatment used are without heating and by heating. Growth and development of plantletswas observed in this study. The results showed that the heating treatment of explants onions cv. Bima Brebes, hasnot reduced the percentage of plantlets infected with the virus. Explants (1/3 bulb) and (Shoot tip), have explantsgrowth above 50%. Generally the smaller explants the higher percentage of abnormal plantlets. Culturecontamination is generally caused by bacteria and fungi that are carried away from explants (endogenous). Thetreatment of heating the onion explants material visually had no effect on the percentage of growth and percentageof contaminated culture.Keywords: red onion, Allium ascolonicum L, heating treatment, source of explant


2018 ◽  
Vol 10 (3) ◽  
pp. 484-490 ◽  
Author(s):  
Zulkarnain Zulkarnain ◽  
Neliyati Neliyati ◽  
Eliyanti Eliyanti

Pineapple propagation by lateral shoots, suckers or crowns is often confronted with limited number of regenerated seedlings and high diversity in flowering and fruit formation. In order to solve this problem, this study offer an alternative method by using tissue culture techniques. This study aimed to determine the effect of growth regulators on plantlet regeneration from bud slicing of pineapple cv. Tangkit. Four levels of 2.4-D (0.0, 0.001, 0.01 and 0.1 ppm) in combination with BA (0.0, 0.1, 1.0 and 10.0 ppm) were tested on solid MS medium. Cultures were incubated in total darkness for a week followed by transfer to 16-hour photoperiod. Results showed that explants treated with 2,4-D and/or BA succeeded in regenerating adventitious shoots. Average leaf number did not differ significantly among treatments (P = 0.60). Highest leaf number (2.99 ± 0.23) was obtained on medium with 0.01 ppm 2,4-D without BA, followed by 0.1 ppm 2,4-D without BA (2.85 ± 0.33). Meanwhile, roots were only formed on medium with 0.1 ppm 2.4-D without BA (4.2 ± 0.37 per shoot). Thus, complete plantlets were regenerated only on medium supplemented with 0.1 ppm 2,4-D without BA. The growth of plantlets was relatively uniform, and plantlet acclimatization succeeded 100% on Jiffy pots. The finding of optimum concentration of 2.4-D and BA in this study is important to develop standard protocol for in vitro propagation of pineapple cv. Tangkit. Thus, the benefit of producing seeds in large quantities and relatively uniform in growth is made possible through tissue culture technique.


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