scholarly journals Characterization of Wolbachia Host Cell Range via the In Vitro Establishment of Infections

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.

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.


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.


Author(s):  
Ata Rofita Wasiati ◽  
Ika Afifah Nugraheni ◽  
Yuli Setiawati

Orchids of the Vanda helvola genus are extensively grown as ornamental plants. The method of propagation of plant seeds in tissue culture is an alternative that may be employed in the provision of a short period. Murashige and Skoog (MS) media and activated charcoal can optimize orchid cultivation utilizing tissue culture techniques. One of the keys to success in tissue culture is the application of natural types of growing regulatory substances (ZPT) at the proper concentration, which can lower the cost of orchid tissue culture.The objective of this study was to see how a combination of Murashige and Skoog (MS) media and activated charcoal influenced the Vanda helvola orchid subculture in vitro. The researchers employed a Complete Randomized Design (RAL) with four treatments and three repeats: negative control (K), ¼ MS (K1), ½ MS (K2), and 100% MS (K3) (K3). The observed parameters include the percentage of live explant, number of roots, and height of orchid plants.   The results showed that the explant had a 100% life percentage of over 21 HST and had no noticeable influence on the number of roots and the plant's height.


2021 ◽  
Vol 13 (1) ◽  
pp. 97
Author(s):  
Muhammad Aris ◽  
Fatma Muchdar ◽  
Rusmawati Labenua

HighlightThe best salinity for the thallus growth of K. alvarezii is 32 mg/LThe salinity indicates the osmotic balance of K. alvarezii seaweedThis osmoregulation process affects the nutrient absorption of K. alvarezii seaweedThis study aims to develop the availibility of superior seeds of K. alvareziiAbstract Kappaphycus alvarezii is one of the leading commodities in Indonesian waters. Demand for this commodity is quite high, as reflected in the increasing volume of exports each year. Fulfillment of these demands is obtained from the production of cultivation. Generally farmers get natural seaweed seedlings, namely cuttings from existing seaweed. The continuous use of seeds from nature can cause deterioration in the quality and quantity. Handling the problem of quality deterioration from seaweed seeds originating from nature, can be overcome by multiplying the seeds through tissue culture methods in vitro. In term of tissue culture techniques method, the most important thing to note is environmental parameters. An environment parameter that changes suddenly such as salinity can inhibit the growth of seaweed. Thus, this work is attampting the different salinity treatment on seaweed explants K. alvarezii. This study aims at determining (weight) the explants of K. alvarezii with a comparison of the different salinity levels in the in vitro tissue culture method. The method used in this study was a completely randomized design (CRD) with the different salinity treatments namely 30, 31, 32, 33, and 34 ppt. The results showed that the different salinities influenced the growth rate of K. alvarezii seaweed explants with the best explant growth at the salinity of 31 ppt, while the lowest growth value was obtained at 34 ppt


2021 ◽  
Vol 21 (1) ◽  
pp. 54-57
Author(s):  
Dyah Nuning Erawati ◽  
Yusriatul Mawaddah ◽  
Siti Humaida ◽  
Irma Wardati

Vanilla has a potential to be developed through tissue culture techniques to anticipate the limitations of the parent plant as a source of planting material. The in vitro propagation ability of vanilla shoots needs to be controlled with the regulation of Kinetin and Benzyl Amino Purines. The interests of this study are 1) analysis of the response of vanilla explants at several Kinetin concentrations; 2) analysis of the response of vanilla explants at several concentrations of BAP and 3) analysis of the interaction of Kinetin and BAP on the response of vanilla explants to form shoot multiplication. The research was conducted at the Tissue Culture Laboratory Politeknik Negeri Jember from June to December 2020 using a factorial Completely Randomized Design (CRD). Factor 1 was the Kinetin concentration of 0.0, 1.0, 2.0 mg.L-1 and the second factor was the concentration of BAP 0.5, 1.5, 2.5 mg.L-1. The results proved that the fastest shoot multiplication occurred on MS medium + Kinetin 2 mg.L-1 with a mean of 8.7 days after inoculation. The mean number of shoots was 7.6 shoots/explant with the highest average wet weight of 0.9 grams/explant at the addition of BAP 1.5 mg. L-1 at measurement 70 days after inoculation.


Jurnal BIOMA ◽  
2014 ◽  
Vol 10 (1) ◽  
pp. 34
Author(s):  
Febrina Ariyanti ◽  
Christiani Tumilisar ◽  
Rossa Yunita

Abstract Cashew (Anacardium occidentale L.) is a plant with high economic value. Conventional propagation of this plant still has obstacles, so an alternative techniques using tissue culture could be tried. One of the factors that determine the success of tissue culture techniques is the type and concentration of growth regulators was used. Growth regulator which have effect on shoot elongation is a cytokinin and gibberellin, this research tried to investigate the influence of combination cytokinin and gibberelin on in vitro shoot elongation of cashew. This research was conducted at BB-Biogen, Bogor on June-November 2010. The method in this research was to design experimental method with completely randomized design. The result was cytokinin could increase the length of shoots and quantity of shoots very well until 4 cm and quantity of shoot for 5. With the most effective cytokinin is zeatin of 5 mg/l.   Key words: Anacardium occidentale L., cytokinin, elongation of shoots, gibberelin


2021 ◽  
Author(s):  
Priyanka Bijalwan ◽  
Shilpa .

In vitro culture of plant cells/tissues is now routine using a range of explant types from many of the important vegetable and fruit crops. Successful technologies include isolation, culture of tissues, cells, protoplasts, organs, embryos, ovules, anthers and microspores and regeneration from them of complete plantlets. The development of plant tissue culture technology represents one of the most exciting advances in plant sciences. For example, the prospect of being able to introduce, develop, produce, transfer and conserve the existing gene pool of plant sciences by using tissue culture methods opens up new opportunities for researches and entrepreneurs. The term plant tissue culture should denote in vitro cultivation of plant cells or tissues in an unorganized mass, i.e., callus culture. Plant tissue culture techniques, in combination with recombinant DNA technology, are the essential requirements for the development of transgenic plants. However, culture techniques like anther/pollen/ovule culture, meristem culture can themselves be utilized for crop improvement or may serve as an aid to conventional breeding. In recent, isolated microspore culture has developed as a breeding tool and an experimental system for various genetic manipulations. The inherent potentiality of a plant cell to give rise to a whole plant, a capacity which is often retained even after a cell has undergone final differentiation in the plant body, is described as ‘cellular totipotency’. On the other hand, production of virus-free plants via meristem culture can reduce losses caused by phyto-pathogens. Embryo culture has many potential uses ranging from overcoming seed dormancy to facilitation of inter-specific hybridization. Protoplast fusion technique can be used for the transfer of cytoplasmic male sterility from one species to another in a short period of time. In cabbage, male sterile cybrids are being utilized by seed companies to produce hybrid seeds on commercial scale and at competitive rates. Plant tissue culture and cell culture are providing useful methods for germplasm storage either by low temperature storage of organized tissue, or cryopreservation of cell or embryo culture.


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