Distribution of polyprenol and dolichol in oil palms from Pisifera parents and mature plants from tissue culture propagation

Kartika EBA, Siregar LAM, Basyuni M. 2021. Distribution of polyprenol and dolichol in oil palms from Pisifera parents and mature plants from tissue culture propagation. Biodiversitas 22: 3423-3436. Oil palm tissue culture is carried out through indirect embryogenesis, which causes somaclonal diversity to occur at the in vitro propagation stage, especially in the callus growth phase. In the cells of all living organisms can be found a group of polyisoprenoid compounds. This study aims to determine variations in oil palm plants resulting from tissue culture based on the presence of polyisoprenoid compounds. Oil palm leaf samples (Elaeis guineensis Jacq.) were collected from the plantation of PT. Socfin Indonesia, North Sumatra, Indonesia. Sample extraction, saponification, isolation of polyisoprenoid compounds, and two-dimensional thin-layer chromatography analysis were carried out to obtain data related to total lipids, polyprenol and dolichol profile, and Carbon (C) chain length polyprenol and dolichol from oil palm leaves of Pisifera parents and their propagated derivatives by indirect embryogenesis. The results showed that the amount of polyisoprenoid mother S24 was 2.41 mg/g higher than that of the parentS8 at 2.17 mg/g dry weight. Total polyisoprenoid fromS8 plants in vitro ranged from 0.71 - 8.53 mg/g dry weight, with the lowest total polyisoprenoid found atS894/22 and the highest was found atS893/2. While for total polyisoprenoid from plant tissue culture of Pisifera S24 ranged from 0.73 - 8.05 mg/g dry weight, with the lowest total polyisoprenoid found at S24H14 and the highest was found at S24H16. The parent plants of Pisifera S8 and S24, as well as plants resulting from tissue culture, were categorized as having lipid pattern type II, which showed a balanced distribution of polyprenol with dolichol. The longest carbon chain was found in vitro plantsS8 93/4 ranged from C50-C110, while the shortest was found in plants produced in vitro S24H7 starting from C45-C55. There were variations in the carbon chain length of polyprenol and dolichol in the leaf samples derived from in vitro propagation of the Pisifera S8 and S24 parents.

BREVIPEDICELLUS and ERECTA mediate expression ofAtPRX17in preventing Arabidopsis callus retardation and browning

ABSTRACTEfficient in vitro callus generation is fundamental to tissue culture propagation, a process required for plant regeneration and transgenic breeding for desired phenotypes. Identifying genes and regulatory elements that prevent callus retardation and browning is essential to facilitate the development of vitro callus systems. Here we show thatBREVIPEDICELLUS(BP) andERECTA(ER) pathways inArabidopsiscallus are converged to prevent callus browning and positively regulate an isoperoxidase gene AtPRX17expression in the rapid growth callus. Loss of functions in bothBPandERresulted in markedly increasing callus browning. Transgenic lines withpro35S::AtPRX17in thebp-5 er105double mutant background fully rescued this phenotypic abnormality. Using plantin vitroDNA-binding assays, we observed that BP protein bound directly to the upstream sequence ofAtPRX17to promote its transcription during callus growth. ER is a universally presenting factor required for cell proliferation and growth, we show thatERpositively regulates expression of a transcription factorWRKY6, which also directly binds to an additional site of the AtPRX17promoter for its high expression. Our data reveals an important molecular mechanism in regulating expression of peroxidase isozyme to reduce Arabidopsis callus browning.HighlightBREVIPEDICELLUSandERECTAare involved in regulating Arabidopsis callus browning by controlling expression ofAtPRX17.

Restoring the natural tropism of AAV2 vectors for human liver

Recent clinical successes in gene therapy applications have intensified interest in using adeno-associated viruses (AAVs) as vectors for therapeutic gene delivery. Although prototypical AAV2 shows robust in vitro transduction of human hepatocyte–derived cell lines, it has not translated into an effective vector for liver-directed gene therapy in vivo. This is consistent with observations made in Fah−/−/Rag2−/−/Il2rg−/− (FRG) mice with humanized livers, showing that AAV2 functions poorly in this xenograft model. Here, we derived naturally hepatotropic AAV capsid sequences from primary human liver samples. We demonstrated that capsid mutations, likely acquired as an unintentional consequence of tissue culture propagation, attenuated the intrinsic human hepatic tropism of natural AAV2 and related human liver AAV isolates. These mutations resulted in amino acid changes that increased binding to heparan sulfate proteoglycan (HSPG), which has been regarded as the primary cellular receptor mediating AAV2 infection of human hepatocytes. Propagation of natural AAV variants in vitro showed tissue culture adaptation with resulting loss of tropism for human hepatocytes. In vivo readaptation of the prototypical AAV2 in FRG mice with a humanized liver resulted in restoration of the intrinsic hepatic tropism of AAV2 through decreased binding to HSPG. Our results challenge the notion that high affinity for HSPG is essential for AAV2 entry into human hepatocytes and suggest that natural AAV capsids of human liver origin are likely to be more effective for liver-targeted gene therapy applications than culture-adapted AAV2.

PERBANYAKAN NILAM (Pogostemon cablin Benth) MENGGUNAKAN MEDIA DASAR ALTERNATIF SECARA IN VITRO In vitro multiplication of Patchouli uses alternative primary medium

<p align="center">Benih sangat menentukan dalam keberhasilan usahatani nilam. Perbanyakan benih secara konvensional vegetatif sangat mudah menularkan penyakit dan membutuhkan waktu yang relatif lama, seperti benih nilam yang diperbanyak selama ini dengan setek. Cara ini memiliki kendala, yang diharapkan dapat diatasi dengan teknik kultur jaringan. Media merupakan faktor utama dalam perbanyakan kultur jaringan. Perbanyakan dan perkembangbiakan tanaman nilam dengan metode kultur jaringan secara umum sudah dapat dilakukan tetapi untuk keberhasilannya sangat tergantung pada jenis media, terutama bila ditinjau dari sisi ekonomi. Karena aplikasi teknologi kultur jaringan untuk tanaman nilam masih dirasakan mahal. Tulisan ini mengulas tentang penggunaan media untuk menekan biaya media kultur jaringan yaitu dengan menggantikan media dasar MS (Murashige-Skoog), ZPT dan vitamin dengan media dasar alternatif dan air kelapa 10%. Air kelapa merupakan salah satu diantara beberapa persenyawaan kompleks alamiah yang sering digunakan dalam kultur jaringan. Sedangkan media dasar alternatif berupa pupuk daun dapat berfungsi sebagai penyedia unsur hara makro mikro dengan komposisi N:P:K (20:20:20). Hal ini untuk mengatasi permasalahan, agar media kultur jaringan menjadi relatif murah, dan harga jual benih lebih terjangkau. Air kelapa yang digunakan berasal dari kelapa hijau yang dicirikan dengan volume air masih memenuhi buah dan keadaan endosperm (daging kelapa) yang belum menebal. Tetapi meski bahan alternatif ini sudah banyak digunakan untuk media pengganti kultur jaringan karena relatif mudah tersedia, murah, menghasilkan benih seragam dan sehat, ternyata belum dapat menunjukkan hasil yang setara dibandingkan dengan penggunaan media MS dalam perbanyakan tunas nilam secara kultur jaringan. Oleh karena itu berbagai penelitian perbanyakan tanaman nilam dengan berbagai metode kultur jaringan agar menghasilkan benih yang murah, sehat, seragam dan dalam jumlah besar masih perlu terus diupayakan.</p><p> </p><p align="center">ABSTRACT</p><p> The quality of seeds are very important in patchoulli cultivation. Cutting multiplication seeds are usually easy in transmitting diseases and relatively need a long time to grow. So far patchouli seeds obtained conventionally with cutting has some constraints, hence tissue culture techniques becomes the solution once. The success of propagation and breeding of plants with tissue culture methods in patchouly is already conducted but is still expensive to be implemented. The paper review patchouli tissue culture propagation by replacing basic media MS (murashige-skoog, Growth Regulating Substances (GRS) and vitamine with alternate basic medium and 10% coconut water Coconut water is one of several natural complex compounds that are often used in tissue culture. The alternative medium as leaf fertilizer can serve as <span style="text-decoration: line-through;">a</span> micro and macro nutrient provider with composition N: P: K (20:20:20). Hopefully, It could become the solution to make tissue culture of patchoulli seeds cheaper and more available. Actually, eventhough the overall substitution of MS medium with full alternative media has already used in limited areas, it has not able yet showing equal results with the use of basic medium MS media in tissue culture patchouli multiplication. Therefore, the researches on patchouli tissue culture should be continued to achieve the huge number, healthy, unity, and unexpensive seeds.</p><p> </p>

Virulensi Isolat Fusarium oxysporum f. sp. cubense VCG 01213/16 pada Pisang Barangan dari Varietas Pisang dan Lokasi yang Berbeda

Analisis genetik isolat-isolat cendawan Fusarium oxysporum f. sp. cubense (Foc) VCG 01213/16 penyebab penyakit layu pada tanaman pisang menunjukkan adanya keragaman yang nyata. Penelitian bertujuan mempelajari keragaman virulensi isolat-isolat yang terkelompok dalam VCG 01213/16, berasal dari berbagai daerah dan varietas pisang yang berbeda. Penelitian dilakukan di Laboratorium Penyakit dan Rumah Kasa Balai Penelitian Tanaman Buah Tropika (Balitbu Tropika) Solok, dari bulan Maret sampai dengan Juni 2009. Rancangan yang digunakan ialah acak kelompok dengan 10 perlakuan dan tiga ulangan, masing-masing perlakuan terdiri atas 10 tanaman. Perlakuan terdiri atas 10 isolat Foc VCG 01213/16 yang berasal dari varietas pisang dan lokasi berbeda. Tanaman uji ialah benih pisang Barangan hasil perbanyakan kultur jaringan. Hasil penelitian menunjukkan bahwa terdapat keragaman virulensi 10 isolat Foc VCG 01213/16 yang dinilai dari perbedaan masa inkubasi, persentase serangan, dan indeks keparahan penyakit pada bonggol dan daun pisang Barangan. Sembilan isolat Foc yang diuji mempunyai virulensi yang tinggi. Masa inkubasi berkisar antara 13,98 dan 16,80 hari, persentase serangan 93,33-100%, dan indeks keparahan penyakit pada bonggol dan daun masing-masing berkisar 3,46-5,35 dan 4,68-5,41. Isolat Foc VCG 01213/16 yang berasal dari Jabung-Lampung Timur dan diisolasi dari pisang varietas Ambon Kuning (isolat F) menunjukkan virulensi yang relatif lebih rendah dibanding sembilan isolat Foc lainnya dengan masa inkubasi 30,27 hari, indeks keparahan penyakit pada bonggol dan daun masing-masing sebesar 2,14 dan 3,76. Hasil penelitian ini bermanfaat dalam memberikan informasi tentang biologi F. oxysporum f. sp. cubense sebagai dasar untuk penyusunan teknik pengendalian yang tepat.<br /><br /><br />Genetic analysis of isolates of the Fusarium oxysporum f. sp. cubense (Foc) that are grouped in VCG 01213/16, as the causal agent of wilt disease in banana plants showed a considerable variation. This research aimed to study the variation in virulence of isolates that are grouped in VCG 01213/16 from different varieties of banana and regions. The study was conducted in the Protection Laboratory and the Screenhouse of Indonesian Tropical Fruit Research Institute (ITFRI) Solok, from March to June 2009. A randomized block design was used in this research with 10 treatments and three replications. Each treatments consisted of 10 banana plants. The treatment was 10 Foc isolates belonging to VCG 01213/16 originating from different varieties of banana and locations. Barangan plantlets produced from tissue culture propagation were used as the planting material. The results showed that there were high variations in virulence among 10 Foc isolates in VCG 01213/16 based on variables of the incubation period, percentage of wilt, and disease severity index on corm and leaves of Barangan variety. Nine of the 10 Foc isolates tested were highly virulent isolates. The incubation period ranged from 13.98 to 16.80 days, the percentage of wilt from 93.33 to 100%, and the disease severity index of corm and leaves ranged from 3.46 to 5.35 and from 4.68 to 5.41, respectively. The Foc VCG 01213/16 isolates originated from Jabung, East Lampung and from Ambon Kuning variety (isolate F) shown relatively low virulence than others isolates that the incubation period was 30.27 days and the disease severity index on the corm and leaves was 2.14 and 3.76, respectively. This result provides useful information on biology of F. oxysprum f. sp. cubense to find out the best control method of the pathogen.<br /><br />