Tissue Culture Laboratory Research Articles

Formation of potato micro tubers (Solanum tuberosum L.) by using BAP and coconut water in the in vitro culture

The use of BAP and coconut water can support the process of forming potato micro tubers in vitro. The research was conducted at the laboratory of tissue culture, Trial Garden of Vegetable Research Center, Berastagi, North Sumatera. This study used a completely randomized 5×4 factorial design. The first factor was BAP concentration at five levels of concentration, i.e 0; 2.5; 5.0; 7.5 and 10 mg/L and the second factor was coconut water concentration at four levels of concentration, i.e 0; 75; 150 and 225 mL/L. The results showed that the best potato explant growth was found at a combination of 0 mg/L of BAP and 0 mg/L of coconut water. The time of potato micro tubers and the fastest number of potato micro tubers were found in the treatment combination of 5 mg/L BAP concentration and 75 mL/L coconut water of coconut water.

Resistance detection of blackjack to ALS inhibitors by in vitro plant growth method

Background: The resistance of weeds to herbicides is a problem that has been increasingly studied because of its great importance in agriculture and rapid growth of this phenomenon worldwide. The quick detection of resistant plants is extremely important for resistance management. Weed resistance to herbicides can be detected through field, greenhouse and laboratory tests. A fast way of detecting resistance is using tissue culture method, where is possible to use resistant plant clones and obtain results faster than whole-plant method in greenhouse. Objective: The aims of the research were to detect the resistance of blackjack to herbicides inhibitors of the acetolactate synthase (ALS), addition in vitro cultivation; determine the herbicide concentration of imazethapyr to 95% control of the susceptible plant population; evaluate explants growth; and, determine the herbicide concentration added to the culture medium where is possible to distinguish a resistant from a susceptible plant. Methods: Experiments were carried out in greenhouse and tissue culture laboratory. Screening was performed to select resistant and susceptible biotypes, tests for specie establishment and dose response curves in vitro were made. Results: The herbicide concentration added to the culture medium that provided 95% susceptible biotype control and efficiently differentiated susceptible biotype from resistant one was 0.6μM of imazethapyr. Conclusions: This method helps recommendations of weed management and provides a quick decision to alternative control of this specie, thus avoiding major damage to the crops.

Somatic embryogenesis on irradiated callus of garlic (Allium Sativum L.)

The research was done at the Tissue Culture Laboratory of Center for Application of Isotope and Radiation Technology, Jakarta. Callus derived from shoot tip of garlic cloves was exposed to gamma rays, then cut into sized of 1 mm3 and cultured in Gamborg medium enriched with plant growth regulators (PGRs) depends on the treatment. The PGRs applied are. The experiment was arranged in a randomized factorial design with 2 factors and 3 replications. The first factor was the doses of gamma rays (0, 10 and 20 Gy) and the second factor was different concentrations of PGRs (untreated, 1 and 2 ppm of thidiazuron (TDZ) and Zeatin). The result indicated that the application of 2 ppm TDZ or Zeatin promoted development on irradiated callus at the dose 10 Gy. The highest number of the shoot and the longest shoot were obtained from non-irradiated callus grown on medium enriched with 2 ppm TDZ, followed by 1 and 2 ppm Zeatin, 1 ppm TDZ and without PGRs. Irradiated callus at the dose 10 Gy grown on medium with 2 ppm TDZ was able to form a shoot, while other irradiated callusat the dose 29 Gy were not able to form the shoot.

Induction and Growth of Endosperm Cali of Rimau Gerga Lebong (RGL) Citrus on Several Media Compositions

Rimau Gerga Lebong (RGL) variety is one of the main orange fruit commodities in Lebong Regency of Bengkulu Province, which has a competitive advantage and has good market potential. However, high number of seed characteristic makes this orange fruit becomes less popular. Triploid genotype formation through endosperm culture in vitro is an alternative solution to develop seedless orange fruit. The objective of this study was to determine the best composition medium for callus induction of endosperm of RGL orange seeds as the foremost step of in vitro triploid plant development. The research was conducted from August 2017 until December 2018 at the Plant Tissue Culture Laboratory, Department of Agronomy, Faculty of Agriculture, University of Bengkulu. The experiment was arranged in a completely randomized design, with eight treatment combinations i.e. G1 (MT + 5 ppm BAP + 2 ppm 2,4-D), G2 (MT + 5 ppm BAP + 2 ppm 2,4-D + 500 ppm CH), G3 (MT + 5 ppm BAP + 2 ppm 2,4-D + 500 ppm + 0.5 ppm Kinetin), G4 (MT + 5 ppm BAP + 2 ppm 2,4-D + 500 ppm CH + 0.5 ppm to 500 ppm Kinetin + ME) , G5 (MS + 5 ppm BAP + 2 ppm 2,4-D), G6 (MS + 5 ppm BAP + 2 ppm 2,4-D + 500 ppm CH), G7 (MS + 5 ppm BAP + 2 ppm 2,4- D + 500 ppm CH + 0.5 ppm Kinetin), G8 (MS + 5 ppm BAP + 2 ppm 2,4-D + 500 ppm CH + 0.5 ppm to 500 ppm Kinetin + ME), with three replications. The experimental unit was five culture bottles containing three explants per bottle. Observations were conducted on days to callus formation, rate of callus formation, callus weight, callus diameter, callus color and texture. The results showed that media of MS + 5 ppm BAP + 2 ppm 2,4-D + 500 ppm CH was the best media for callus induction, and MT + 5 ppm BAP + 2 ppm 2,4-D + 500 ppm CH was the best media for callus development.Keyword: RGL citrus, calli, endosperm, kinetin, BAP

Accuracy of Fuzzy Logic-based Contamination Grading System on Abaca Tissue Culture

This study aimed to support the coalition for abaca rehabilitation program of the different cooperating agencies of Southern Leyte with the use of intelligent systems in the grading of abaca tissue culture contamination. As the Southern Leyte State University Tissue Culture Laboratory has since used manual grading of contaminated specimen, the use of image acquisition technology can lessen the human effort involved in this activity. Fuzzy logic was used in the grading of the contamination. There were five inputs for the inference engine, namely red, green, blue, whitish and brownish. There were 62 rules identified covering the different combinations, and the triangular-shaped membership function was used for all the membership types of the inputs. Based on the result of the testing, it had an accuracy rate of 82.00% for the binary contamination in 50 sample specimens, and an over-all accuracy of 80.33% in multiclass contamination grading. The system could not match the precision of the human expert, but the model is comparable to that of the expected actual result, while minimizing human intervention in grading the contamination of tissue cultured abaca.

An Assessment of the Viability of Producing True-to-type, Virus-free, Tissue-cultured Garlic in the Philippines

Considering that the Philippines’ garlic production cannot cope up with the local demand requirements, this supply gap presents itself as a bright spot in garlic farming. As the demand for garlic far outpaces production levels which remain low due to poor yields exacerbated by traditional growing practices of garlic farmers, there is a need to demonstrate whether producing true-to-type, virus-free, tissue-cultured garlic propagation is the answer to bridge such a supply gap. This study, which is part of a larger research study focused on looking into the technology utilization of producing true-to-type, virus-free, tissue-cultured (TC) garlic planting materials for farmers, aimed to evaluate the feasibility of producing in vitro bulblets from multiplied shoots and bulb production under greenhouse and field conditions so that better quality and high volumes of garlic planting materials may be available to garlic farmers. Data gathering was done through face-to-face interviews with tissue culture laboratory personnel, farmers and municipal agriculture officers and from secondary and online sources. Using comparative analysis with traditional garlic production methods, cost and return, benefit-cost ratio and NPV, it was revealed that TC garlic planting materials are more costly and beyond the reach of farmers. Moreover, current traditional garlic practices are more profitable compared to TC garlic production (whether as G0 in vitro bulblets from multiple shoots or as G2 bulblets). Though the sales potential of farmers using TC planting materials is about three times more than that of traditional farmers, their net income is only half of that of traditional farmers because challenges remain in terms of bringing the production cost of TC planting materials down in view of lack of economies of scale. It is imperative that continuing improvements in research laboratory protocols be done to bring down the gestation period of TC planting materials from 3 years to one year and address the high contamination rates in the production process. In addition, economies of scale must be considered in choosing at which production level to operate.

Callus Induction of Aerides odorata Lour. by Adding 2,4 Dichlorophenoxyacetic Acid (2,4-D)

The research about callus induction of Aerides odorata L. by adding 2,4 Dichlorophenoxyacetic Acid (2,4-D), has been done from August to October 2018 in Plant Physiology and Tissue Culture Laboratory, Department of Biology, Faculty of Mathematics and Natural Science, Andalas University, Padang. The aim of this research was found the effective consentration of 2,4-D to induce somatic embryo of A.odorata. The research used Completely Randomized Design (CRD) with 5 treatments and 6 replications. The treatments were: 0 mg/L 2,4-D; 0,25 mg/L 2,4-D; 0,5 mg/L 2,4-D; 0,75 mg/L 2,4-D; 1 mg/L 2,4-D. The result showed that the treatmeant were able induction callus of A.odorata, with compact until the friable texture, color of the resulting callus is yellowish green and greenish yellow. 2,4-D 1 mg/L was the best concentration to increase fresh weight of callus.

Pengaruh Pemberian BAP dan NAA terhadap Pertumbuhan Krisan (Chrysanthemum morifolium, Ram.) dalam Kultur Jaringan

The objective of this study was to evaluate the effect of combination benzyl amino purine(BAP) and naphtalene acetid acid (NAA) on tissue culture of Chrysanthemum. Theexperiment was conducted at The Tissue Culture Laboratory, Agriculture Faculty,Stiper Agriculture Institute. The Completely Randomized Design was applied in thisexperiment, consisting of two factors; those were BAP and NAA application. Each factorconsisted of four treatments. Each combination of treatment was carried out with ninereplications. The conclusion from this study were: Application of higher BAP and NAAconcentration increased budding of explant, whereas application of higher NAA thanBAP concentration increased the growth of bud and leaf number, also increased rootingof explant.

Optimizing Quality of White Oyster Mushroom Seeds Through Plant Propagation

Quality seeds determine the productivity of white oyster mushrooms (Pleurotus ostreatus). The problem is the interaction between F0 seed inoculation (agar medium) and F1 seed media. The research objective of the two-factor correlation results produces high-quality seeds and optimal growth power. The research conducted at the Tissue Culture Laboratory of the Faculty of Agriculture, Merdeka University, Madiun. Two-factor experiments in the Factorial Randomized Design repeated three times: A (inoculation of F0 seed media to make A1 potato extract and media to extract coconut water A2). B (F1 media growing grain) consists of B1 corn, B2 rice grain, B3 red beans, B4 soybeans, B5 corn seeds and rice grains, B6 red beans and soybeans, B7 corn and soybeans. Anova statistical analysis continued Duncan test 5%. Significant interaction at the time of mycelium appearance, mycelium growth rate and the percentage of mycelium spread ages 7, 14 and 21 The highest time of mycelium appearance (1.8 DAP) in the combination of A1 B7 (F0 seed media for potato extract with growing medium of corn seeds and soybean seeds) and A2 B3 (media F0 seedlings to extract coconut water on rice grain growing media). Increasing speed for mycelia is the highest combination of treatment A1 B2 (F0 seed media for potato extract with rice grains) = 2.2 HIS. Percentage of 100% mycelium spread at age 21 DAP in combination A1 B1 (A: F0 seed potato extract with B2 growing medium of corn seeds). A2 (F0 seed extract of coconut water) inoculated on the medium of corn seeds and corn seeds plus other seeds of rice, red beans and soybeans provide significant results.

Effect of Rhustox on Micropropagation of Scoparia dulcis L. through Leaf Explants Culture

Aim: To investigate the effect of Rhus toxicodendron (30CH) along with different compositions of phytohormones (Auxin and Cytokinin) on the basis of growth and multiplication of explants under optimum temperature under in-vitro conditions.
 Study Design: To establish and design the standard protocol for the in-vitro propagation through leaf explant of Scoparia dulcis under stress of phytohormones and homeopathic medicine Rhus toxicodendron (30CH).
 Place and Duration of Study: The plant materials were procured from the Herbal Botanical Garden Patna Science College, Department of Botany, Patna University, Patna, Bihar. The experimental part was carried out in Plant Tissue Culture Laboratory, between December 2017 to August 2018 in Department of Botany P.U. Patna.
 Methodlogy: The sterilized leaf explants were inoculated into MS media fortified with different phytohormones (Auxin and Cytokinin) and Rhus tox(30CH) under aseptic environmental conditions for the growth and development of callus, embryoids etc.
 Result: The explants in MS medium supplemented with auxins phytohormones and Rhus tox(30CH) exhibited that IAA (0.10 to 2.0 mg/l) and BAP (0.10 to 2.5 mg/l) induces green and compact calli. Whereas at 0.30mg/l of IAA and 0.50 mg/l BAP induced brown friable calli. 2,4-D (1.5 mg/l) and Kinetin (1.5-6.5mg/l) concentrations induced brown and friable calli. Rhus tox(30CH) (100 µl/100 ml) enhances proliferation with 2,4-D and Kinetin (1.5/1.5 mg/l.).
 Conclusion: After 42 days of culture initiation and establishment the callus was 520.0±1.12 mg in the mixture of 2,4-D and Kinetin (1.5 mg/l) in Rhus tox free medium. Whereas weight of callus were found to be 1092±0.74 mg after 42 days in the same medium of 2, 4-D and Kinetin (1.5/5.5 mg/l) supplemented with Rhus tox (100 µl/100 ml). Hence, the investigation proponded that the Rhus tox (CH30) has increased the rate of callus development and plantlet regeneration.

In-vitro Callus Induction of Durian (Durio zibethinus Murr.) Leaves Using Kinetin and 2,4-D (Dichlorophenoxyacetic acid)

Durian (Durio zibethinus Murr.) is a tropical fruit grown in Southeast Asia and it has high nutritional and economic values and highly appreciated by consumers in Indonesia. Unfortunately, there are several obstacles in planting durian in Indonesia, specifically in Aceh. One of the abstacles is the scarcity of superior durian plants. Generally, plant conservations done conventionally through vegetative propagation. However, this technique has disadvantage such as harming the initial plants. Therefore, plant culture tissue has been used as a modern technique to develop durian plants. The objective of this research was to evaluate the effect of kinetin and 2,4 D on development of durian leaf explants through in-vitro bioassay. The research was conducted at Tissue Culture Laboratory, Faculty of Agriculture, Malikussaleh University from January to April 2019 using Completely Randomized Design (CRD) Factorial with 2 factors observed. The first factor was Kinetin (K): 0.0, 0.1 and 0.5 ppm. The second factor was auxin 2,4 D: 0.0, 0.5 and 1.0 ppm. The results showed that the in-vitro application of kinetin affected the induction of leaf callus. The application of 0.5 ppm of kinetin gave the best result compared to others. The in-vitro application of 2,4 D 0.5 ppm also possessed the best result compared to others. There was no interaction beween in-vitro application of kinetin and 2,4 D on the growth of durian leaf callus at all parameters observed.

RESPONSE OF TURMERIC SHOOT EXPLANT AFTER CYTOKININ WITH IN VITRO

This study was aimed at determining the response of turmeric shoot explants after the provision of various types and concentrations of cytokinins in vitro. This experiment was conducted at the Tissue Culture Laboratory, Faculty of Agriculture, Padjadjaran University from January to April 2018. The data were analyzed using T-Test. The experimental method used in this research was Completely Randomized Design (CRD). Explant planting was carried out in Laminar Air Flow. The experiment consisted of 7 treatments consisting of 4 replications and each test consisted of 4 units. Observation of this experiment was carried out for 12 MST. The main observations were made on the data that were tested statistically namely the percentage of explant growing shoots, percentage of explant growing roots, shoot height, number of tuns, number of roots and root length. The treatments consisted of Control, 2.5 mg L-1 BAP, 5 mg L-1 BAP, 0.5 mg L-1 TDZ, 1 mg L-1 TDZ, 0.01 mg L-1 Zeatin and 0.1 mg L Zeatin -1. The results show that the treatment of 1 mg L-1 TDZ shows the best response to the growth of turmeric explants by increasing the number of turmeric shoot explants than the others.RESPONS EKSPLAN TUNAS KUNYIT SETELAH SITOKININ SECARA IN VITROTujuan dari penelitian ini yaitu untuk melihat respons eksplan tunas kunyit terhadap pemberian berbagai jenis dan konsentrasi sitokinin secara in vitro. Percobaan ini dilakukan di Laboratorium Kultur Jaringan, Fakultas Pertanian, Universitas Padjadjaran dari bulan Januari sampai April 2018. Hasil percobaan dianalisis dengan Sample T-Test. Metode percobaan yang digunakan dalam penelitian ini yaitu Rancangan Acak Lengkap (RAL). Penanaman eksplan dilakukan di dalam Laminar Air Flow. Percobaan terdiri dari 7 perlakuan sebanyak 4 ulangan dan setiap ulangan terdiri dari 4 unit. Pengamatan percobaan ini dilakukan selama 12 MST. Pengamatan utama dilakukan terhadap data-data yang diuji secara statistik yakni persentase eksplan tumbuh tunas, persentse eksplan tumbuh akar, tinggi tunas, jumlah tunas, jumlah akar dan panjang akar. Perlakuan terdiri dari Kontrol; 2,5 mg L-1 BAP; 5 mg L-1 BAP; 0,5 mg L-1 TDZ; 1 mg L-1 TDZ; 0,01 mg L-1 Zeatin; dan 0,1 mg L-1 Zeatin. Hasil penelitian menunjukkan bahwa perlakuan 1 mg L-1 TDZ menunjukkan respons yang lebih baik terhadap pertumbuhan eksplan kunyit dengan meningkatkan jumlah tunas eksplan tanaman kunyit daripada yang lainnya.

Effect of light specters (red and blue) on two-potato varieties tissue (spunta and Agria)

This study was carried out in plant tissue culture laboratory which belong to bio technology research center in order to test the effect of raid and blue spectral light colors and white lighting as a control and white fluorescent lamps as a second control on plant growth stages while two potato variety ( Spunta , Agria) were used in this experiment . potato plant samples were collected and cultured in special jars contain MS media after being sterilized by Clorox solution in order to avoid the any contamination risk , while the culture stage of the explant started in sterilized condition in sited the hood cabinet, wherever the culture stage completed the cultured plants were placed in the growth room under controlled condition with raid , blue and normal light condition. The obtained results showed that the wait normal color treatment was significantly higher than the other two treatments in plant length factor for the tow used potato variety, while for the number of leaves factor the results showed that the normal light color treatment was significantly higher than the other two treatments in plant number of leaves factor followed by raid color treatment in the two used variety Spunta and Agria while the Raid color treatment scored average of leaves (23.9, 23.3 ) leaf respectively.

Effect of adding growth regulators benzyl adenine (BA) and kinetin (Kin) on production of potato (spunta) micro tuber

This experiment was carried out in plant tissue culture laboratory a part of bio technology research center (BTRC) , while the main aim for this experiment was to investigate the effects of using plant growth regulators on potato micro tuber formation for Sponta potato variety throw plant tissue culture method , In the beginning micro propagation operations were done to the target potato variety in order to obtain enough number of plants samples in free contamination MS media , while different concentrations of BA and Kin plant growth regulators ( 0 , 1 , 2 , 3 mg/l ) were used in this experiment , the plant growth regulators BA ,Kin were added to MS media contain 60g/l sucrose during micro tuber formation stage . the results of this study showed that the treatment of (5mg/l Kin ) obtained the best results and this treatment gave the highest tuber production compared with other treatments , furthermore this treatment ( 5mg/l Kin) gave the highest number of produced potato tuber and the highest weight of produced tuber compared with other used treatments , while the average number of produced micro tubers arrived to 4.9 tuber for each plant and the average weight for obtained potato tuber arrived to 0.84 g which was higher than other weight of micro tuber produced from the other treatments .

Effect of Sucrose on in vitro Bud Multiplication of Torch Ginger (Etlingera elatior)

Torch Ginger (Etlingera elatior (Jack) R. M. Smith) is plant material that can be used as a medicine, food, cosmetics, and bio-pesticides. The existence of torch ginger in nature is becoming more limited due to the utilization of health care services mainly as a medicine continues to increase. In conventionally, plant propagation of torch ginger is derived from a rhizome, but seedlings produced tend to have the level of low proliferation and susceptible to infections of soil pathogens. This research aimed to know the influence of sucrose and BAP concentration on bud growth of torch ginger in in vitro. The research was conducted in the Tissue Culture Laboratory Universitas Islam Negeri Sunan Gunung Djati Bandung. Research method used was a descriptive design with experiment treatments i.e. sucrose (20 g/L, 40 g/L, and 60 g/L) and BAP (0 mg/L and 1 mg/L). The result showed that the use of sucrose 40 g/L + BAP 1 mg/L gave the best response to the shoot growth on 6 WAI and capable to produce green shoots on 8 WAI.

Effect of growth regulators on in vitro micropropagation of blue honeysuckle (Lonicera caerulea L.)

The experiment was carried out in the Tissue Culture Laboratory of the College of Agriculture, University of Karbala to investigate the effect of growth regulators in micropropagation of the blue honeysuckle in vitro. The branches were cultivated by shoot tips in MS medium with five concentrations of each BA and Kin @ 0, 1.0, 1.5, 2.0 and 2.5 mg/l and each concentration was mixed with IAA @ 0.2 mg/l at multiplication stage. At rooting stage, independent experiments were conducted using four concentrations of each IBA and NAA @ 0, 0.5, 1.0 and 1.5 mg/l and each concentration was mixed with steady concentration of 0.2 mg/l of BA. The results indicated that marked differences were observed in plant growth characters under different concentrations of cytokines used. The cytokine concentration of 2.5 mg/l produced significantly higher number of branches, length of branch, and wet and dry weight of vegetative parts compared to control. As for the effect of cytokines type, the BA showed its ’superiority over other cytokines in terms of production of higher number of branches, length of branch, and wet and dry weight of vegetative parts. It was also observed that the auxin types and their concentrations significantly affected the number and length of roots and their wet and dry weights. The IBA concentration of 1.5 mg/l achieved significantly higher number of branches, length of branch, and wet and dry weight of vegetative parts of blue honeysuckle compared to control where no auxin was applied.

PENGARUH KONSENTRASI BAYCLIN PADA PENCUCIAN II DAN BAP PADA MEDIA MS TERHADAP PERTUMBUHAN EKSPLAN TANAMAN PISANG KLUTUK (Musa paradisiaca. L) SECARA IN VITRO

This research aims to examie the effect of bayclin concentration on washing II and BAP in MS media on eksplan growth of klutuk banana in vitro and was conducted at the Tissue Culture Laboratory Lempes Pekanbaru from April to August 2016. The research used a completely randomized design (CRD) with 3 factorials and 3 replications. The first factor was the concentration of washing eksplan bayclin on phase II which consisted of four levels: K0 (concentration of 0%), K1 (concentration of 10%), K2 (concentration of 20%), and K3 (concentration of 30%). The second factor was the concentration of BAP contained on media MS consisted of four levels: T0 (BAP concentration 0 mg/liter), T1 (BAP concentration of 0.1 mg/liter), T2 (BAP concentration of 1.0 mg/liter) and T3 (BAP concentration of 10.0 mg/liter). The parameters observed were percentage of explants contaminated, age appears buds, the number of shoots/explant, length of shoots/explant, number of roots/explant (root), root length/explant and percentage of explant life. The results showed that effect of concentration on the washing bayclin II and BAP in MS Media on growth eksplan of klutuk banana in-vitro was the best treatment K2T2 with 33.33% of explants contaminated, age of appears buds 2,67 weeks after planting, the number of shoots 2,67/explant, length of shoots 5,50 cm/explant, number of roots 2.67/explant, root length 12,08 cm/explant and 58.33% of explants life.

In Vitro Screening Ketahanan Galur Padi (Oryza Sativa) B7 Hasil Rakitan Politeknik Negeri Lampung Terhadap Keracunan Besi (Fe)

This research aimed to invstigate the response of paddy culture (B7 strain) assembled by Lampung State Polytechnic to the iron toxicity tolerance. The research was done at Plant Tissue Culture Laboratory, Lampung State Polytechnic, from July to September 2019. Treatments were single arranged in a completely randomized design with three replications. The treatment tried was five levels of Fe concentrations (5,6 ppm 28 ppm, 56 ppm, 84 ppm, 112 ppm, and control). Each replication consisted of three culture bottle containing one explant. The homogenity of data was tested using Barlett test. If the assumption were fulfilled then analysis of variance is executed using STATISTIX 10, and then followed by the Honest Significant Difference (HSD) test in 5% alpha for mean separation and RPA analysis. The result of this research showed that the B7 strain has tolerance to iron toxicity until 56 ppm of Fe concentration, it can be concluded from the PAR value of its strain (>0,50). Meanwhile in 84 and 112 ppm of Fe concentration, the RPA value of B7 starin (<0,50), and it is indicate that its strain is sensitive.Â

Influence of biotic elicitor Aspergillus niger on salicylic acid products in callus cultures of Calendula officinalis L. plant

The study was carried out in the college of education for pure sciences / department of biology / plant tissue and cell culture laboratory to study the effect of biotic elictors Aspergillus niger in the production of salicylic acid in callus cultures of Calendula officinalis L. plant. the results showed the best induction of the explants, which included the leaves and cotyledon obtained from culturing on the solidified MS medium supplemented with 2.0 mg.L1- of 2,4- Dichlorophenoxy acetic acid (2,4-D) interacted with 0.5 mg.L−1 Kinetin, which reached the highest fresh weight of callus was 2.2350 and 2.7125 g, respectively, after four weeks culture. The callus induction from the leaves its compact, in contrast to the callus induced by the cotyledon, which is characterized its friable.

EFFECT OF GROWTH REGULATOR NAA AND IBA APPLICATIONS ON TOTAL PHENOLIC AND FLAVONOID COMPOUNDS EXTRACTED FROM IN VITRO PRODUCED CALLUS OF CHICORY PLANT (Cichorium intybus L.)

This research study was carried out in the plant tissue culture laboratory of the Agricultural Botany Department, Faculty of Agriculture, Ain Shams University, Shoubra El-khaima, Cairo, Egypt. Experiments were executed for the duration of two consecutive years 2017 and 2018 on chicory plant. Chicory (Cichorium intybus L.), which belongs to Asteraceae family, is considered as an important medicinal plant due to the presence of many bioactive substances such flavonoids, phenolic compounds, alkaloids, steroids, terpenoids, including( coumarines, cichoriin, esculetin, inulin, sesquiterpene lactones, chicoric acid, caffeic acid and some vitamins). In this research in vitro experiments were carried out using full strength Murashige and Skoog basal medium (MS) supplemented with different combinations of two plant growth regulators; Indole-3-butyric acid (IBA) including two concentrations (0.5 – 2.0 mg/l) and Naphthalene acetic acid (NAA) comprising four concentrations (0.5 – 2.0 – 3.0 – 5.0 mg/l). An abaxially (lower side) leaf explants (square pieces 0.5 × 0.5 cm) which were taken from 20 days old aseptic chicory seedlings were inoculated to (MS) surface. Initially, chicory seeds were aseptically germinated on half-strength MS medium, after surface sterilization by 70 % (v/v) ethanol for 60 seconds then soaking in 10 % Clorox (0.5% sodium hypochlorite NaOCl) for 10 min to produce the aseptic chicory seedlings which were the source of true leaf explants used in this research study. Total phenolic compounds and flavonoids content were extracted from six-week C. intybus friable callus produced under both light and dark in vitro culture conditions inside a growth chamber incubation room where temperature was adjusted at 25oC ±1. Total phenolic compounds and flavonoids were determined by spectrophotometric methods. The highest values for their contents were from chicory calli when MS callus induction medium was supplemented with 2 mg/l NAA under total dark condition when compared with the other remaining growth regulator treatment combinations and alternative light regime conditions.