575.222.7:581.1 doi: 10.15407/biotech8.01.019
Institute of Cell Biology and Genetic Engineering of the National Academy of Sciences of Ukraine,
Received 12.01.2015
The review explores some of the recent advances and the author's own researchs concerning biotech-nological approaches for Agrobacterium tumefaciens- and A. rhizo.genes-mediated transformation of Compositae family plants. This paper reviews the results of genetic transformation of Compositae plants, including edible (Cichorium intybus, Lactuca sativa), oil (Helianthus annuus), decorative (Gerbera hyb-rida), medical (Bidens pilosa, Artemisia annua, Artemisia vulgaris, Calendula officinalis, Withania somnifera etc.) plant species. Some Compositae genetic engineering areas are considered including creation of plants, resistant to pests, diseases and herbicides, to the effect of abiotic stress factors as well as plants with altered phenotype. The article also presents the data on the development of biotechnology for Compositae plants Cynara cardunculus, Arnica montana, Cichorium intybus, Artemisia annua "hairy" roots construction.
Key words: Compositae, Agrobacterium tumerfaciens, Agrobacterium rhizogenes, "hairy" roots.
Eighties of the 20th century were the beginning of active development and widespread adoption of genetically engineered plants. The development of methods of gene cloning and methods of transferring of genetic information into a plant genome served as background and stimulation of this process. The study concerning Genetic Engineering is dedicated to basic research of the functioning of the transferred genes and practical use of new biotechnological approaches. First efforts were directed at developing of biotechnology to obtain plants with novel features, such as resistance to pests, diseases and to abiotic stresses. These plants should also provide an increased synthesis of biologically active compounds and recombinant proteins. Transformation using Agrobacterium tumefacience and A. rhizogenes is a simple and effective method of construction of biotechnological plants (plants with transformed genome). These soil pathogenic bacteria are able to transfer part of their DNA, Ti- or Ri-plasmids to plant genome
[1-3]. The using of foreign genes transferring technology into plant DNA is possible due to natural properties of phytopathogenic bacteria and progress in cloning technology. Transformation using A. tumefaciens and development of methods of shoot regeneration for certain species allowed to obtain transgenic plants for a relatively short period of time. This method is usually used to create new forms of crops resistant to diseases and pests, plants which synthesize recombinant proteins (called "edible" vaccines), decorative plants with new phenotype. The special attention has to be paid to the use of bacteria A. rhizogenes for genetic transformation and construction of "hairy" roots which are characterized by genetic stability [4] and grow on medium without growth regulators. Growing such roots in bioreactors is not expensive and can be used to produce bioactive compounds of plant origin [5-9]. Nowadays, the system of genetic transformation and creation of transgenic roots for a large number of plants have been developed. The possibility to increase the
content of bioactive compounds which are naturally synthesized in an original plants has been shown [10-16].
Agrobacterium-mediated transformation is also used to create Compositae plants with a new characteristics. Compositae family includes about 2,000 species.
Among them there are medicinal (e. g., Arnica, Inula, Calendula, Tragopogon, Tussilago, Artemisia, Sylibium, Stevia, Echinacea), oil (Helianthus), decorative (Gerbera, Cosmos, Dahlia, Bellis, Chrysanthemum, Echinacea) and edible (Lactuca, Cichorium, Cynara) plants. Compositae family plants are used in traditional medicine, they reveal antiradical and antioxidant activity [17-19], synthesize compounds with anti-inflammatory [20], hepatoprotective [21-23], cytotoxic [24-26], antiparasitic [27], antimicrobial [28] and immunomodulatory [29] properties. Many genetic engineering experiments on the Compositae plants were carried out to improve their resistance to diseases, pests and abiotic stress factors, to increase the content of important compounds, to select the plants with altered phenotype. In addition, the study was designed to create plants that synthesize recombinant proteins with therapeutic properties. The following table provides information on the most significant studies on Agrobacterium-mediated transformation of Compositae family plants.
Optimization of conditions
of genetic transformation
The first investigations on genetic transformation of Compositae plants were dated in the late eighties of the 20th century. They were aimed at fundamental development of the transformation system using bacteria A. tumefaciens and A. rhizogenes. It is known that the efficiency of transformation depends on the complex of factors which are associated with both virulent activity of agrobacteria strains and morphological, physiological and characteristics of transformed plants. Selection of transgenic plants (correct choice of selective and reporter genes) is an important step of transformation. Therefore, the definition of critical» i.e. principal for plant species parameters is essential to obtain transgenic plants or roots. Considering all above mentioned factors, it is necessary to define the optimal type of explants, transforming conditions, using different strains of agrobacteria, reporter and selective genes etc. The greatest amount of research has
been aimed at developing of transformation protocols for the plant species, which are very important for agriculture and medicine such as Lactuca sativa, Artemisia annua, Cichorium intybus and Helianthus annuus. Lettuce and chicory plants are used for food; chicory, thanks its compounds with therapeutic properties, is used as a base for certain medicines. That is why these species cultivated in vitro have been studied in detail. For example, to obtain transgenic plants of lettuce the effect of plant genotype, the concentration of bacteria in suspension for transformation and a period of transformation on the efficiency of transformation were determined [61]. Optimal conditions for transformation of lettuce in particular the time of cocultivation with bacteria were defined in [72].
Efficiency of mannose phosphate isome-rase, neomycin phosphate transferase II, beta glucuronidase selective genes and reporter gus gene usage to select L. sativa and C. intybus transgenic plants was investigated [39, 40, 72].
The influence of plant genotype on the regenerative ability of three C. intybus (Witloof, Melci, Hera) cultivars and obtaining of transgenic plants using A. tumefaciens were studied [38]. We obtained L. sativa and C. intybus var foliosum transgenic roots using A. rhizogenes-mediated transformation method (wild strain A4) and defined the conditions of shoot regeneration from "hairy" roots. Formation of transgenic shoots on the roots of chicory was occurred spontaneously in culture medium without hormones under lighting conditions whereas lettuce regeneration was possible only after cultivation on the medium which included growth regulators such as kinetin and a-naphtylacetic acid (Figure). The rolB gene of A. rhizogenes was detected in regenerated plants as well as in "hairy" roots.
Artemisia is another species of Compositae family. It includes herbaceous plants which are common in Europe, America, Central Asia and North Africa.
The common wormwood (A. vulgaris L.), annual mugwort (A. annua), absinthium (A. absinthium), tarragon (A. dracunculus) and other species belong to this family. These plants are used in official and alternative medicine, so there is a great interest in these species, especially in China and India, where this plant material is widely used in medical practice. A. rhizogenes and A. tumefaciens were used to perform genetic transformation of Artemisia sp. As it was shown in [49] A. annua was transformed by the A. rhizogenes LBA 9402 strain, transgenic roots were obtained and the
Agrobacterium-mediated transformation of plants of Compositae family
Species Usage The way of transformation The result of transformation References
Arnica montana L. pharmaceutical A. rhizogenes Transgenic roots were obtained [30]
Arnica montana L. pharmaceutical A. rhizogenes The light influence on the synthesis of thymol derivates in transgenic roots was investigated [11]
Dahlia pinnata decorative . tumefaciens Transformation conditions were optimized [31]
Gerbera hybrida decorative . tumefaciens Plants with altered pigmentation were obtained [32]
Gerbera hybrida decorative . tumefaciens Transformation conditions were optimized [33], [34]
Gerbera jemosonii decorative . tumefaciens Transformation conditions were optimized [35]
Gerbera hybrida decorative . tumefaciens Gene of resistance to tomato spotted wilt virus was transferred into plants [36]
Cynara carduncuus edible . tumefaciens gus-positive callus was obtained [37]
Cichorium intybus edible pharmaceutical . tumefaciens Transformation conditions were optimized [38], [39], [40]
Cichorium intybus edible pharmaceutical Agrobacterium rhizogenes Regenerated plants with early flowering were obtained [41]
Cichorium intybus edible pharmaceutical Agrobacterium rhizogenes The influence of transformation on phenotype altering was defined [42], [43]
Cichorium intybus edible pharmaceutical Agrobacterium tumefaciens Plants resistant to chlorine sulfon were obtained [44]
Cichorium intybus edible pharmaceutical Agrobacterium tumefaciens The plants resistant to salinity after their transformation by gene AtNHXl gene were obtained [45]
Artemisia annua pharmaceutical Agrobacterium rhizogenes The conditions of transformation were optimized [46]
Artemisia annua pharmaceutical Agrobacterium rhizogenes The conditions of cultivation of transgenic roots to increase arte-misinin synthesis were defined [47]
Artemisia annua pharmaceutical Agrobacterium rhizogenes The conditions of transformation were optimized, the efficiency of usage of different strains of bacteria for transformation was defined. [48], [49], [50]
Artemisia annua pharmaceutical Agrobacterium tumefaciens The plants with altered flowering time were obtained [51]
Artemisia annua pharmaceutical Agrobacterium tumefaciens An effective transformation protocol was developed [52]
Artemisia dubia and Artemisia indica pharmaceutical Agrobacterium rhizogenes The conditions of growing and synthesis of artemisinin in transgenic roots were investigated [53]
Artemisia annua pharmaceutical Agrobacterium tumefaciens The conditions of transformation were optimized with the usage of the different strains of agrobacteria [54], [55]
Species Usage The way of transformation The result of transformation References
Artemisia vulgaris pharmaceutical Agrobacterium rhizogenes The conditions of transformation were optimized with the usage of the different strains, different expiants and under different cultivation conditions [56]
Artemisia absinthium pharmaceutical Agrobacterium rhizogenes The culture of transgenic roots was obtained, the production of secondary metabolites was defined [57]
Taraxacum platycarpum pharmaceutical Agrobacterium rhizogenes The changes of morphology after transformation were described [58]
Inula helenium edible Agrobacterium rhizogenes The conditions of transformation were optimized [59]
Lactuca sativa edible Agrobacterium tumefaciens Expression of ABF3 gene of Arabidopsis plants increased resistance to drought and cold [60]
Lactuca sativa edible Agrobacterium tumefaciens The conditions of transformation were optimized [61]
Lactuca sativa edible Agrobacterium tumefaciens ipt-gene was used under the control of SAG12 promoter to prevent aging leaves [62]
Lactuca sativa edible Agrobacterium tumefaciens The genes that determine resistance to destruction by insects (pta, ct, cgrp) were transferred [63], [64]
Lactuca sativa edible The plants resistant to the herbicide (bar-gene) were obtained [65]
Lactuca sativa edible Agrobacterium tumefaciens The increase of resistance to abiotic stresses (drought, low temperature) using ABF3, P5CS genes was determined [66], [67], [68]
Lactuca sativa edible Agrobacterium tumefaciens The transgenic seed was obtained in vitro [69]
Lactuca sativa edible Agrobacterium tumefaciens Plant genes that ensure the resistance to phytovirus (coat protein genes) were integrated into plant genome [70], [71]
Lactuca sativa edible Agrobacterium tumefaciens Efficiency of transformation and methods of selection of transgenic plants (genes of mannose phosphate isomerase, neomycin phosphate transferase II) were defined [72]
Centaurea montana pharmaceutical Agrobacterium tumefaciens The conditions of transformation were optimized [73]
Bidens pilosa pharmaceutical Agrobacterium tumefaciens The conditions of transformation were optimized, the gene of halcon synthase gene was transferred [74]
Tanacetum cine-rariifolium Agrobacterium tumefaciens The conditions of transformation were optimized (selection in the presence of hygromycin) [75]
Helianthus annuus x Helianthus tuberosus oil Agrobacterium rhizogenes The method for hybrids rooting was developed [76]
Helianthus annuus oil Agrobacterium tumefaciens The efficiency of transformation with the usage of different genotypes and types of explant was compared [77], [78] [79], [80]
Species Usage The way of transformation The result of transformation References
Helianthus annuus oil Agrobacterium tumefaciens The conditions of transformation were optimized, the efficiency of usage of different reporter and selective genes (gus, gfp, mgf5, nptll) was defined [81], [82], [83], [84], [85], [86], [87] [88], [89]
Helianthus annuus oil Agrobacterium tumefaciens Resistance to phosphinotricin [90]
Helianthus annuus oil Agrobacterium tumefaciens Plants resistant to white rot were obtained [91]
Helianthus annuus oil Agrobacterium rhizogenes Transgenic roots were obtained [92]
Helianthus annuus oil Agrobacterium tumefaciens Plants with increased synthesis of L-proline and resistant to abiotic stress were obtained [93]
possibility of spontaneous regeneration of shoots from the roots on non-hormonal medium MC was revealed. The efficiency of usage of A. rhizogenes A4, 15834, K599, LBA 9402, 9365, 9340 for Artemisia plants transformation was estimated [48]. The authors also determine the efficiency of acetosyringon addition to the culture medium during cultivation of agrobacteria or to the medium during explants co-cultivation with bacterial suspension. The genetic transformation of A. annua using A. rhizogenes LBA 9402 was carried out, the transfer of TL-DNA (rol gene) of agrobacterial plasmid was confirmed by PCR analysis [50]. Since the purpose of this study was to create transgenic roots which are characterized by rapid growth and high accumulation of artemisinin, the authors found out the influence of temperature, pH, medium composition and carbon source on "hairy" roots growth. The effect of agrobacteria strains used (A. rhizogenes LBA 9402, LBA 920, LBA 301, MTCC 532, NRRL B193 and A4) on the efficiency of A. annua transformation was studied. The most suitable type of explant (leaf, stem, petiole) was also revealed [56]. Artemisia dubia, A. indica, A. absinthium, A. vulgaris "hairy" root cultures were obtained using A. rhizogenes-mediated transformation method [53, 56, 57]. We firstly transformed A. tilesii Ledeb (Aleutian wormwood) plants using efficient transformation protocol. We obtained the transgenic roots with the frequency 100% (Figure) under optimized conditions of co-cultivation with A. rhizogenes 4. Transgenic A. annua plants were obtained using A. tumefaciens. The protocol of A. annua transformation [52] was optimized. It was shown that this process was effected by
A. tumefaciens strain, plant genotype, usage of acetosyringon, the period of co-cultivation with bacteria etc. The similar studies with the usage of different strains of agrobacteria (LBA4404, GV1301, AGL1, EHA105) and different conditions of transformation (age of explants, the method of inoculation of bacteria, duration and conditions of co-cultivation of explants with bacteria) were carried out in [54] and [55]. Nowadays number of Compositae plant species which are used for genetic transformation significantly expands. It includes for instance Cynara cardunculus [37], Gerbera jemosonii [35], Inula helenium [59], Centaurea montana [73], Bidens pilosa [74], Tanacetum cinerariifolium [75], Dahlia pinnata [31]. The progressive interest in the new species is associated with their ability to synthesize commercial biologically active compounds. So, artichoke can synthesize the compounds possessing antioxidant and hepatoprotective activity. The gus-positive (30%) transgenic callus was obtained using Agrobacterium tumefaciens-mediated transformation and transgenic Cynara cardunculus roots with 45% frequency were obtained using A. rhizogenes-mediated transformation [37]. Two strains of A. rhizogenes (AR15834 A4) were used to obtain transgenic roots of Inula helenium. The influence of explant type on the transformation process was defined and the transgenic roots from leaf and stem explants of different age were obtained [59]. Transgenic Centaurea montana plants were constructed using two A. tumefaciens strains, AGL1 and EHA105. However, the efficiency of the transformation was low (1.8% from 990 explants) [73]. In our opinion the great attention may be paid to the usage of Bidens
pilosa (spanish needle), the tropical plant which has a well-known Ukrainian relative Bidens tripartita. 21 lines of regenerated plants from 1,373 explants were obtained as the result of the recent research [74] using Agrobacterium-mediated transformation (vector pCHS with chs chalcone synthase Petunia gene and selective nptll gene). At 15 plants two transferred genes were detected.
Creation of plants resistant to biotic and abiotic stress factors
Among the Compositae plants there are many species which have economic value and are grown as oil and green cultures. Helianthus annuus is an important oil plant, Lactuca sativa and Cichorium intybus are salad plants. Higher yields of these crops can be achieved by creating plants which are resistant to insects, viral and bacterial diseases, abiotic stress (drought, soil salinity etc). Creation of plants resistant to insects, phytopathogenic microorganisms and abiotic stress factors is aimed at increasing yield of crops and efficiency of agriculture. Since ninetieth of the 20th century the great amount of investigations has been dedicated to the development of the system for genetic transformation of sunflower. This oilseed is widely cultivated in the countries of European community, Russian Federation and Argentina. Ukraine is the leader of sunflower growing and the part of Ukrainian production in the global harvest has been increased during the last decade. The interest in sunflower biotechnology is amplified. The optimal conditions of co-cultivation with agrobacteria and selection in the presence of kanamycin after transformation was studied. The efficiency of usage of acetosyringon and some reporter genes (gus, gfp, mgf5) was defined. Transformation efficiency using mature and immature embryos, leaves, varieties or hybrids of H. annuus was compared [83-90]. The content of culture medium was optimized and the influence of genotype on the efficiency of transformation of Helianthus annuus was determined [78]. Transformation using A. rhizogenes was performed to provide better rooting of hybrids [76]. The investigated technological schemes served as a base to obtain crops with valuable properties. Thus, sunflower plants resistant to phytopathogenic fungi Sclerotinia sclerotiorum were obtained using A. tumefaciens-mediated transformation [91]. H. annuus plants resistant to the herbicide Basta at a concentration of 3 l/ha
by transferring bar gene were obtained [90]. Ukrainian researchers analyzed the feasibility of using double-stranded RNA suppressor gene of prolin dehydrogenase (based on Arabidopsis ProDHl gene) to increase the resistance of sunflower plants to stress factors such as water deficiency and salinity. Transgenic H. annuus plants with a high content of free L-proline under the stress conditions were constructed and a reduction of content of this compound during recovery plants after stress was noted. So these transgenic plants possessed the better adaptation abiliy to stress conditions [93].
Chicory and lettuce are plants grown in different regions (Europe, Asia and America). Some directions of improving the quality of these plants were aimed at obtaining plants resistant to stress factors. As it was reported [45] the chicory plants resistant to salinity were constructed by A. tumefaciens--mediated transformation. Transferring of AtNHXl gene in chicory genome not only increased the tolerance of plants to salinity, but also reduced the damage of cell membranes induced by a high salt content. Transferring of P5CS gene, which encodes the enzyme delta-pirrolin 5-carboxyl-synthase involved in the synthesis of proline, made it possible to increase the resistance of lettuce plants to abiotic stress [67]. It is known that abscisic acid acts in the response of plants to the effect of stress factors. Transgenic lettuce plants resistant to drought and cold stress were obtained using ABF3 gene, cloned from Arabidopsis [68]. Viral coat protein genes in two modifications were transferred into Lactuca sativa by A. tumefaciens--mediated transformation and as a result LBVaV virus was not detected in the obtained virus-infected plants.
It was established that CP gene in antisense orientation permit a resistance to phytovirus for one line of plants [71]. Transferring bar gene to L. sativa gave the possibility to obtain plants resistant to the herbicide [65]. The possibility of an efficient transferring Pta gene (Pinellia ternata Agglutinin) into the lettuce plants have been reported, however the researchers didn't define the resistance of transgenic plants to insects [63].
Construction of plants
with altered phenotype
Genetic transformation can lead to a changes in plant phenotype. Such changes were observed at chicory plants which were transformed by A. rhizogenes [41]. Regenerated plants formed flower-bearing stems in the
^ansgenic roots:
Cichorium intybus (a), Lactuca sativa (c) and regeneration of plants from these roots (b, d)
first year of growth, although these plants are perennial. This peculiarity of genetic transformation and transformation with the usage of specific genes made it possible to isolate new forms of decorative plants. So, transformation of decorative Gerbera jemosonii plants with the usage of two genes which are responsible for the color of the flowers - iris-dfr and petunia-f3'5'h have led to the change of flowers pigmentation [35]. Transferring chalcone synthase gene antisense cDNA to gerbera plants provided the suppression of the anthocyanin synthesis and has led to change of pigmentation of some regenerated plants [33]. Although the number of studies concerning the usage of Compositae plants is rather limited, the biotechnological approaches to obtain new forms are promising because of wide usage of decorative plants e.g. Gerbera and significant progress which was made in biotechnological methods to change phenotype of decorative plants [94].
The Compositae family includes numerous species of interest for biotechnological research. However, only some of them are using now in genetic engineering to obtain plants with valuable economic features. There are no investigations in genetic transformation
of such plants as Ligularia thomsonii, Xanthium stramarium, Scorzonera undulata, Senecio erucifolius, Tussilago farfara. Some publications dedicated to biotechnological approaches to create transgenic Stevia rebaudiana [95], Bidenspilosa [74], Echinacea purpurea [96, 97] plants is limited. At the same time there is a group of plants that have practical value. Thus, Asteriscus plants synthesize essential oils, which possess antioxidant properties; Vernonia condensata, Scorzonera undulata, Bidens pilosa are the source of antioxidants; some species synthesize flavonoids, phenolic compounds, sesquiterpene lactones; fructose containing compounds with wide spectrum of biological activity, in particular hepatoprotective, prebiotic, antidiabetic, immunomodulatory activity etc.
So, a genetic transformation of new plant species to improve their properties (resistance to stress factors, plants with altered phenotype) as well as creation of plants producing recombinant proteins besides naturally synthesized compounds is a preferable direction of practical usage of plant biotechnological approaches.
1. Chilton M. D, Tepfer D. A., Petit A., Chantal David, Francine Casse-Delbart, Jacques Tempe. Agrobacterium rhizogenes inserts T-DNA into the genomes of the host-plant root cells. Nature. 1982, V. 295, P. 432-434. doi:10.1038/295432a0.
2. Tepfer D. Transformation of several species of higher plants by agrobacterium rhizogenes: Sexual transmission of the transformed genotype and phenotype. Cell. 1984, 37 (3), 959-967. PMID:6744417.
3. Matzke A. J. M.,Chilton M. D. Site-specific insertion of genes into T-DNA of the Agrobacterium tumor-inducing plasmid: an approach to genetic engineering of higher plant cells. J. Mol. Appl. Genet., 1981, 1 (1), 39-49. PMID:6955419.
4. Baiza A. M., Quiroz-Moreno A., Ruiz J. A., Loyola-Vargas V. M. Genetic stability of hairy root cultures of Datura stramonium. Plant Cell Tiss. Org. 1999, 59 (1), 9-17. doi: 10.1023/A:1006398727508.
5. GeorgievM.,PavlovA.I.,Bley T. Hairy root type plant in vitro systems as sources of bioactive substances. Appl. Microbiol. Biotechnol. 2007, 74 (6), 1175-1185. PMID:17294182.
6. Shi H. P., Long Y. Y, Sun T. S, Tsang P. K. E. Induction of hairy roots and plant regeneration from the medicinal plant Pogostemon Cablin. Plant Cell Tiss. Org. 2011, 107 (2), 251-260. doi: 10.1007/s11240-011-9976-9.
7. Mishra B. N, Ranjan R. Growth of hairy-root cultures in various bioreactors for the production of secondary metabolites. Biotechnol. Appl. Biochem. 2008, 49 (1), 1-10. PMID:18086010.
8. Bais H. P., Suresh B., Raghavarao K., Ravishankar G. A. Performance of hairy root cultures of Cichorium intybus L. in bioreactors of different configurations. In Vitro Cell Dev. Biol. Plant. 2002, 38 (6), 573-580. doi:10.1079/IVP2002334.
9. Tang K, Shen Q., Yan T., Fu X. Transgenic approach to increase artemisinin content in Artemisia annua L. Plant Cell Rep. 2014, 33 (4), 605-615. doi: 10.1007/s00299-014-1566-y.
10. Bais H. P., Sudha G., Ravishankar G. A. Enhancement of growth and coumarin production in hairy root cultures of Cichorium intybus, L. cv. Lucknow Local (Witloof Chicory) under the influence of fungal elicitors. J. Biosci. Bioeng. 2000, 90 (6), 640-645. PMID:16232926.
11. Weremczuk-Jezyna I., Kalemba D, Wysokinska H. Constituents of the essential oil from hairy roots and plant roots of Arnica montana. J. Essent. Oil Res. 2011, 23 (1), 9197. doi:10.1080/10412905.2011.9700432.
12. Rahnama H., Hasanloo T., Shams M. R., Sepehrifar R. Silymarin production by hairy root culture of Silybum marianum (L.) Gaertn. Iran. J. Biotechnol. 2008, 6 (1), 113-118.
13. Mahesh A., Jeyachandran R. Agrobacterium rhizogenes-mediated hairy root induction in Taraxacum officinale and analysis of sesquiterpene lactones. Plant Biosystems.
2011, 143 (3), 620-626 doi: 10.1080/ 11263504.2011.584702.
14. Malarz J., Stojakowska A., Kisiel W. Sesquiterpene Lactones in a Hairy Root Culture of Cichorium intybus.Z. Naturforsch. 2002, 57 (11-12), 994-997. PMID:12562083.
15. Samadi A., Carapetian J., Heidari R., Ja-feri M. A. Hassanzaden Gorrttapen Hairy Root Induction in Linum mucronatum ssp. mucronatum, an Anti-Tumor Lignans Producing Plant. Not. Bot. Horti. Agrobo.
2012, 40 (1), 125-131.
16. Bandyopadhyay M., Jha S., Tepfer D. Changes in morphological phenotypes and withanolide composition of Ri-transformed roots of Withania somnifera. Plant Cell Rep. 2007, 26 (5), 599-609. PMID:17103214.
17. Syed Hamid Hussain, Abdul Latif, Cox R. J., Simpson T. J., Mumtaz Ali, Mohammad Arfan, Ghias Uddin. http:// pii/S1874390013001602 - aff0020mailto:j. Phytochemicals from the aerial parts of Ligularia thomsonii and their radical scavenging activity. Phytochem. Lett. 2014, 7 (6), 6-10. doi: 10.1016/j. phytol.2013.09.002.
18. Akbar Esmaeili, Zahra Mousavi, Maryam Shokrollahi, Ali Shafaghat. Antioxidant Activity and Isolation of Luteoline from Centaurea behen L. Grown in Iran. of Chem. 2013,
19. Stojakowska A., Malarz J., Szewczyk A., Kisie W. Caffeic acid derivatives from a hairy root culture of Lactuca virosa. Acta Physiol. Plant. 2012, 34 (1), 291-298. doi:10.1007/ s11738-011-0827-4.
20. Albino de Almeida A. B., Snchez-Hidalgo M., Martn A. R., Luiz-Ferreira A., Trigo J. R., Vilegas W., dos Santos L. C., Souza-Brito A. R., de la Lastra C. A. Anti-inflammatory intestinal activity of Arctium lappa L. (Asteraceae) in TNBS colitis model. J. Ethnopharmacol. 2013, 146 (1), 300-310. doi: 10.1016/j.jep.2012.12.048.
21. Shih-Chang Chien, Young P. H., Yi-Jou Hsu, Chen C. H., Tien Y. J., Shiu S. Y., Li T. H, Yang C. W., Marimuthu P., Tsai L. F., Yang W. C. Anti-diabetic properties of three common Bidens pilosa variants in Taiwan.
science/article/pii/S0031942209002854 -corlPhytochemistry. 2009, 70 (10), 12461254. doi: 10.1016/j.phytochem.
22. Li-Ping Yuan, Fei-Hu Chen. http://www. S0378874108000147 - cor1mailto:cfhchina@, Lu Ling, Dou P. F, Bo H, Zhong M. M., Xia L. J. Protective effects of total flavonoids of Bidens pilosa L. (TFB) on animal liver injury and liver fibrosis. J. Ethnopharmacol. 2008, 116 (3), 539-546. doi: 10.1016/j.jep.2008.01.010.
23. Heibatollah S, Reza N. M., Izadpanah G, Sohailla S. Hepatoprotective effect of Cichorium intybus on CCl4-induced liver damage in rats. Afr. J. Biochem. Res. 2008, 2 (6), 141-144.
24. Kviecinski M. R, Felipe K. B, Schoenfelder T, de Lemos Wiese L. P., Rossi M. H., Gongalez E., Felicio J. D., Filho D. W., Pedrosa R. C. Study of the antitumor potential of Bidens pilosa (Asteraceae) used in Brazilian folk medicine. J. Ethnopharmacol. 2008, 117 (1), 69-75. PMID:18342465.
25. Mohamed Salla, Isabelle Fakhoury, Najat Saliba, Darwiche N., Gali-Muhtasib H. Synergistic anticancer activities of the plant-derived sesquiterpene lactones salograviolide A and iso-seco-tanapartholide. J. Nat. Med. 2013, 67 (3), 468-479. doi: 10.1007/ s11418-012-0703-6.
26. Hughes R., Rowland I. R. Stimulation of apoptosis by two prebiotic Chicory fructans in the rat colon. Carcinogenesis. 2001, 22 (1), 43-47. PMID:11159739.
27. Medjroubi K., Benayache F., Bermejo J. Sesquiterpene lactones from Centaurea musimomum. Antiplasmodial and cytotoxic activities. Fitoterapia. 2006, 76 (7-8), 744776. doi: 10.1016/j.fitote.2005.08.005.
28. Milosevic T., Argyropoulou C., Solujic S., Murat-Spahic D., Skaltsa H. Chemical composition and antimicrobial activity of essential oils from Centaurea pannonica and C. jacea. Nat. Prod. Commun. 2010, 5 (10), 1663-1668. PMID:21121269.
29. Nergard C. S., Diallo D., Michaelsen T. E, Malterud K. E., Kiyohara H., Matsumoto T., Yamada H., Paulsen B. S. Isolation, partial characterisation and immunomodulating activities of polysaccharides from Vernonia kotschyana Sch. Bip. ex Walp. J. Ethnopharmacol. 2004, 91 (1), 141-152. PMID:15036481.
30. Petrova M., Zayova E., Vlahova M. Induction of hairy roots in Arnica montana L. by Agrobacterium rhizogenes. Central Europ. J. Biol. 2013, 8 (5), 470-479. doi:10.2478/ s11535-013-0157-6.
31. Yuko Otani, Dong Poh Chin, Masahiro Mii. Establishment of Agrobacterium-mediated
genetic transformation system in Dahlia. Plant Biotechnol. 2013, 30 (2), 135-139.
32. Elomaa P., Honkanen J., Puska R., Sepp-nen P., Helariutta Y., Mehto M., Kotilai-nen M., Nevalainen L., Teeri T. H. Agrobacterium-Mediated Transfer of Antisense Chalcone Synthase cDNA to Gerbera hybrida Inhibits Flower Pigmentation. Nat. Biotechnol. 1993, 11 (4), 508-511. doi:10.1038/nbt0493-508.
33. Nagaraju V., Srinivast G. S. L., Lakshmi Sita G. Agrobacterium-mediated genetic transformation in Gerbera hybrida. Curr. Sci. 1998, 74 (7), 10-15.
34. Lee Hye-Young, Lee Ki-Jung, Jeon Eun-Hee, J eon Eun-Hee, Shin, Sang-Hyun, Lee Jai-Heon, Kim Doh-Hoon, Chung Dae-Soo, Chung Yong-Mo, Cho Yong-Cho, Kim Jeong-Kook, Chung Young-So. Optimization of Genetic Transformation Conditions for Korean Gerbera Lines. J. Plant Biotechnol. 2006, 33 (1), 49-56. doi: 10.5010/JPB.2006.33.1.049.
35. Hussein G. M., Abu El-Heba G. A., Abdou S. M., Abdallah N. A. Optimization of transient gene expression system in Gerbera jemosonii petals.GM Crops Food. 2013, 4 (1), 50-57. doi: 10.4161/gmcr.23925.
36. Korbin M., Podwyszynska M., Komorowska B., Wawrzynczak D. Transformation of Gerbera plants with Tomato Spotted Wilt virus (TSWV) nucleoprotein gene. Proc. XX EUCARPIA Symp. on New Ornamentals II Eds. J. Van Huylenbroeck. Acta Hort. 572, ISHS. 2002, P. 149-157.
37. Menin B., Comino C., Moglia A., Dolzhenko Y. Setting up of genetic transformation system in globe artichoke. Proceedings of the 54th Italian Society of Agricultural Genetics Annual Congress Matera, Italy, 27/30 September, 2010.
38. Maroufi Asad, Karimi Mansour, Khosro Mehdi Khanlou, Van Bockstaele E., De Loose M. Regeneration ability and genetic transformation of root type chicory (Cichorium intybus var. sativum). Afr. J. Biotechnol. 2012, 11 (56), 11874-11886.
39. Frulleux F., Weyens G., Jacobs M. Agrobacterium tumefaciens-mediated transformation of shoot-buds of chicory. Plant. Cell Tiss. 1997, 50 (2), 107-112. doi: 10.1023/A:1005994711865.
40. Cheng Lin Mei, Cao Qiu Fen, Huang Jing. Establishment of a highly efficient genetic transformation system in Cichorium intybus. Acta Prataculturae Sinica. 2004, 13 (6), 112-116.
41. Bais H. P., Venkatesh R. T., Chandrashekar A., Ravishankar G. A. Agrobacterium rhizogenes-mediated transformation of Witloof chicory in vitro shoot regeneration and induction of flowering. Curr. Sci. 2001, 80 (1), 83-87.
42. Sun L. Y., Gerard T., Charbonnier C., Tepfer D. Modification of phenotype in Belgian endive (Cichorium intybus) through genetic transformation by Agrobacterium rhizogenes: conversion from biennial to annual flowering. Transg. Res. 1991, 1 (1), 14-22. doi:10.1007/BF02512992.
43. Kamada H., Saitou T., Harada H. No requirement of vernalization for flower formation in Ri-transformed Cichorium plants. Plant Tiss. Cult. Lett. 1992, 9 (3), 206-208.
44. Vermeulen A., Vaucheret H., Pautot V., Chupeau Y. Agrobacteriu-mediated transfer of a mutantArabidopsisacetolactatesynthase gene confers resistance to chlorsulfuron in chicory (Cichorium intybus L.). Plant. Cell. Rep. 1992, 11 (5-6), 243-247. doi: 10.1007/ BF00235074.
45. Zhao Yu Wei, Wang Ying Juan, Bu Huai Yu, Hao Jian Guo, Jia Jing Fen. Transformation of Cichorium intybus with the AtNHX1 gene and salinity tolerance of the transformants. Acta Prataculturae Sinica. 2009, 18 (3), 103-109.
46. Mukherjee S., Ghosh B., Jha T. B., Jha S. Genetic transformation of Artemisia annua by Agrobacterium rhizogenes. Ind. J. Exp. Biol. 1995, N 33, P. 868-871.
47. Weathers P. J., Bunk G., McCoy M. C. The effect of phytohormones on growth and artemisinin production in Artemisia annua hairy roots. In Vitro Cell Dev. Biol. Plant. 2005, 41 (1), 47-53. doi: 10.1079/ IVP2004604.
48. Giri A., Ravindra S. T., Dhingra V., Nara-su M. L. Influence of different strains of Agrobacterium rhizogenes on induction of hairy root and artemisinin production in Artemizia annua. Curr. Sci. 2001, 81 (4), 4-25.
49. Banerjee S., Zehra M., Gupta M. M., Kumar S. Agrobacterium rhizogenes-mediated transformation of Artemisia annua: production of transgenic plants. Planta Med. 1997, 63 (5), 467-469. PMID: 17252369.
50. Ahlawat S., Saxena P., Ram M., Pravej Alam, Tazyeen nafis, Anis Mohd, Malik Zainul Abdin. Influence of Agrobacterium rhizogenes on induction of hairy roots for enhanced production of artemisinin in Artemisia annua L. Plants. Afr. J. Biotechnol. 2012, 11 (35), 8684-8691.
51. Wang H., Liu Y., Chong K., Liu B. Y., Ye H. C., Li Z. Q., Yan F., Li G. F. Earlier flowering induced by over-expression of CO gene does not accompany increase of artemisinin biosynthesis in Artemisia annua. Plant Biol. 2007, 9 (3), 442-446. PMID: 17099845.
52. Han J. L, Liu B. Y, Ye H. C., Hong Wang, Zhen-Qiu Li, Guo-Feng Li. Effects of overexpression of the endogenous farnesyl diphosphate synthase on the artemisinin content in Artemisia annua L. Acta Botanica Sinica. 2006, 48 (4), 482-487. doi: 10.1111/ j.1744-7909.2006.00208.
53. Mannan A., Shaheen N, Ar shad W, Qure-shi R. A., Muhammad Zia, Bushra Mirza. Hairy roots induction and artemisinin analysis in Artemisia dubia and Artemisia indica. Afr. J. Biotechnol. 2008, 7 (18), 3288-3292.
54. Alam P., Mohammad A., Ahmad M. M., Khan M. A., Nadeem M, Khan R, Akmal M, Ahlawat S, Abdin M. Z. Efficient method for Agrobacterium mediated transformation of Artemisia annua L. Rec. Path. Biotechnol. 2014, 8 (1), 102-107. PMID: 22642822.
55. Elfahmi, Suhandono S., Chahyadi A. Optimization of genetic transformation of Artemisia annua L. Using Agrobacterium for Artemisinin production. Pharmacogn. Mag. 2014, 10 (37), 176-180. doi: 10.4103/09731296.127372.
56. Sujatha G., Zdravkovic-Korac S., Calic D, Flamin G., Ranjitha Kumari B. D. High-efficiency Agrobacterium rhizogenes-mediated genetic Transformation in Artemisia vulgaris: Hairy root production and Essential oil analysis. Industr. Crops Prod. 2013, V. 44, P. 643-652. doi: 10.1016/j. indcrop.2012.09.007.
57. Nin S., Bennici G., Roselli D., Mariotti D., Schiff S., Magherini R. Agrobacterium-mediated transformation of Artemisia absinthium L. (wormwood) and production of secondary metabolites. Plant Cell Rep. 1997, 16 (10), 725-730. doi: 10.1007/ s002990050310.
58. Lee M. H., Yoon E. S., Jeong J. H., Choi Y. E. Agrobacterium rhizogenes-mediated transformation of Taraxacum platycarpum and changes of morphological characters. Plant Cell Rep. 2004, 22 (11), 822-827. PMID: 14986056.
59. Zahra Shirazi, Khosro Piri, Asghar Mirzaie Asl, Tahereh Hasanloo. Establishment of Inula helenium hairy root culture with the use of Agrobacterium rhizogenes. Int. Res. J. Appl. Basic Sci. 2013, 4 (5), 1034-1038.
60. Enkhchimeg V., Tae W. B., Key Z. R., Soo-Young Kim, Hyo-Yeon Lee. Overexpression of Arabidopsis ABF3 gene enhances tolerance to drought and cold in transgenic lettuce (Lactuca sativa). Plant Cell Tiss. Organ Cult. 2005, 83 (1), 41-50. doi: 10.1007/s11240-005-3800-3.
61. Michelmore R. W., Marsh E., Seely S, Landry B. Transformation of lettuce (Lactuca sativa) mediated by Agrobacterium tumefaciens. Plant Cell Rep. 1987, 6 (6), 439-442. PMID: 24248927.
62. McCabe M. S., Lee C. Garratt, Schepers F., Jordi W. J. R. M., Stoopen G. M., Davelaar E, J. Hans A. van Rhijn, Brian Power J., Davey M. R.. Effects of PSAG12-IPT Gene Expression on Development and Senescence in Transgenic Lettuce. Plant Physiol. 2001, 127 (2), 505-516. PMC125086.
63. Ahmed M. B., Akhter M. S., Hossain M. An Efficient Agrobacterium-mediated Genetic Transformation Method of Lettuce (Lactuca sativa L.) With an Aphidicidal Gene, Pta (Pinellia ternata Agglutinin). Middle-East J. Sci. Res. 2007, 2 (2), 155-160.
64. Valimareanu S. Leaf Disk Transformation of Lactuca sativa Using Agrobacterium tumefaciens. Not. Bot. Hort. Agrobot. Clu.
2010, 38 (3), 181-186.
65. Mohapatra U., McCabe M. S., Power J. B, Schepers F., Van Der Arend A., Davey M. R. Expression of the Bar Gene Confers Herbicide Resistance in Transgenic Lettuce. Transgen. Res. 1999, 8 (1), 33-44. doi: 10.1023/A:1008891216134.
66. Pileggi M., Pereira A. A. M., Silva J. dos Santos, S. Alvim Veiga Pileggi, Verma D. Pal S. An Improved Method for Transformation of Lettuce by Agrobacterium tumefaciens with a Gene that Confers Freezing Resistance. Braz. Arch. Biol. Technol. 2001, 44 (2), 191-196.
67. Pileggi M. Genetic transformation of the lettuce cultivar Grand Rapids (Lactuca sativa L.) by Agrobacterium tumefaciens to improve osmotic stress tolerance. Genet. Mol. Res. 2002, 1 (2), 176.
68. Vanjildorj E., Bae T. W., Riu K. Z., Soo-Young Kim, Hyo-Yeon Lee. Overexpression of Arabidopsis ABF3 gene enhances tolerance to drought and cold in transgenic lettuce (Lactuca sativa). Plant Cell Tiss. Org. Cult. 2005, 83 (1), 41-50. doi: 10.1007/s11240-005-3800-3.
69. Franklin G., Oliveira A. L., Dias, A. C. P. In vitro flowering and viable seed setting of transgenic lettuce cultures. Plant Biotechnol.
2011, 28 (1), 63-68.
70. Dinant S., Maisonneuve B., Albouy J., Chupeau Y., Chupeau M.-Ch, Bellec Y., Gaudefroy F., Kusiak C., Souche S., Robaglia C., Herv Lot. Coat protein gene-mediated protection in Lactuca sativa against lettuce mosaic potyvirus strains. Mol. Breed. 1997, 3 (1), 75-86. doi: 10.1023/A:1009671925550.
71. Yoichi Kawazu, Ryoi Fujiyama, Keita Sugiyama, Takahide Sasaya. A Transgenic Lettuce Line with Resistance to Both Lettuce Big-vein Associated Virus and Mirafiori Lettuce Virus. JASHS. 2006, 131 (6), 760-763.
72. Liu Jingmei, Chen Daming, Chen Hang. Genetic Transformation and Plant Regeneration of Lettuce with Sweet Protein Gene MBLII. Acta Hort. Sin. http://www. shtml2001, http : //www. ahs abstract/abstract 3015. shtml28 http : // shtml abstract3015.shtml(3), 246-250.
73. Abou-Alaiwi W. A., Potlakayala S. D., Goldman S. L., Puthiyaparambil C. Josekutty, Deepkamal N. Karelia, Sairam V. Rudrab-hatla Agrobacterium-mediated transformation of the medicinal plant Centaurea montana. Plant Cell Tiss Org. Cult. 2012, 109 (1), 1-8. doi: 10.1007/s11240-011-0067-8.
74. Chen-Kuen Wang, Shin-Yun Hsu, Po-Yen Chen, Kin-Ying To. Transformation and characterization of transgenic Bidens pilosa L. Plant Cell Tiss. Org. Cult. 2012, 109 (3), 457-464. doi:10.1007/s11240-011-0110-9.
75. Mao J., Cao L. Y., Kong L. F., Maarten A Jongsma science/article/pii/S0304423812005055, Cai-Yun Wang. An Agrobacterium-mediated transformation system of pyrethrum (Tanacetum cinerariifolium) based on leaf explants. Sci.Horticult. 2013, N 150, P. 130134. doi:10.1016/j.scienta.2012.10.019.
76. Prathibha Devi http://www. science/ article/ pii/S0304423801003223 CORR1, Swaroopa Rani. Agrobacterium rhizogenes induced rooting of in vitro regenerated shoots of the hybrid Helianthus annuus x Helianthus tuberosus. Sci. Horticult. 2002, 93 (2), 179-186. doi: 10.1016/S0304-4238(01)00322-3.
77. Grel E., Kazan K. Evaluation of various sunflower (Helianthus annuus L.) genotypes for Agrobacterium tumefaciens-mediated gene transfer. Turk. J. Bot. 1999, N 23, P. 171-177.
78. Haixue Liu, Xiaodong Xie, Shoujun Sun, Wenbi Zhu, Jing Ji, Gang Wang. Optimization of Agrobacterium-mediated transformation of sunflower (Helianthus annuus L.) immature embryos. AJCS, 2011, 5 (12), 1616-1621.
79. Sujatha M., Vijay S., Vasavi S., Reddy P. Veera, Chander Rao S. Agrobacterium-mediated transformation of cotyledons of matureseeds of multiple genotypes of sunflower (Helianthus annuus L.). Plant Cell Tiss. Org. Cult. 2012, 110 (2), 275-287. doi: 10.1007/sll240-012-0149-2.
80. Anitha V., Farzana Jabeen, Ansari N. A., Padma V. Genetic transformation studies in sunflower (Helianthus annuus L.). J. Res. ANGRAU. 2012, 40 (1), 91-93.
81. Knittel N., Gruber V., Hahne G., Lenee P. Transformation of sunflower (Helianthus annuus L.): a reliable protocol. Plant Cell Rep. 1994, 14 (2-3), 81-86. doi: 10.1007/ BF00233766.
82. Burrus M., Molinier J., Himber C., Hunold R., Bronner R., Rousselin P., Hahne G. Agrobacterium-mediated transformation of sunflower (Helianthus annuus) shoot apices: transformation patterns. Mol. Breed. 1996, 2 (4), 329-338. doi:10.1007/BF00437911.
83. Lucas O., Kallerhoff J., Alibert G. Production of stable transgenic sunflowers (Helianthus annuus L.) from wounded immature embryos by particle bombardment and co-cultivation with Agrobacterium tumefaciens. Mol. Breed. 2000, 6 (5), 476-487. doi:10.1023/A:1026583931327.
84. Mller A., Iser M., Hess D. Stable transformation of sunflower (H. annuus L.) using a non-meristematic regeneration protocol and green fluorescent protein as a vital marker. Trans. Res. 2001, 10 (5), 435444. PMID:11708653.
85. Hewezi T., Jardinaud F., Alibert G., Kallerhoff J. A new protocol for efflcient regeneration of a recalcitrant genotype of sunflower (Helianthus annuus L.) by organogenesis induction on splitembryonic axis. Plant Cell Tiss. Org. Cult. 2003, N 73, P. 81-86.
86. Weber S., Friedt W., Landes N., Molinier J., Himber C., Rousselin P., Hahne G., Horn R. Improved Agrobacterium-mediated transformation of sunflower (Helianthus annuus L.): assessment of macerating enzymes and sonication. Plant Cell Rep. 2003, 21 (5), 475482. PMID:12789451.
87. Mohamed S., Boehm R., Binsfeld P. C., Schnabl H. Agrobacterium-mediated transformation of two high oleic sunflower (Helianthus annuus L.) genotypes: assessment and optimizationof important parameters. Helia. 2004, 27 (40), 25-40.
88. Ikeda M., Matsumura M., Kamada H. Suitability of small and branching sunflower varieties for molecular genetic experimentsand their transformation by
Agrobacterium infection. Plant Biotechnol. 2005, 22 (2), 97-104. doi: http://dx.doi. org/10.5511/plantbiotechnology.22.97.
89. Mohamed Sh., Boehm R., Schnabl H. Stable genetic transformation of high oleic Helianthus annuus L. genotypes with high efficiency. Plant Sci. 2006, 171 (5), 546554. doi: 10.1016/j.plantsci.2006.05.012.
90. Neskorodov Y. B., Rakitin A. L., Kamionska-ya A M., Skryabin K. G. Developing phosphinothricin-resistant transgenic sunflower (Helianthus annuus L.) plants. Plant Cell Tiss. Org. Cult. 2010, 100 (1), 6571. doi: 10.1007/s11240-009-9620-0.
91. Sawahel W., Hagran A. Generation of white mold disease-resistant sunflower plants expressing human lysozyme gene. Biol. Plant. 2006, 50 (4), 683-687. doi: 10.1007/ s10535-006-0106-1.
92. Tao Jun, Tan Rufang, Li Ling. Genetic transformation of sunflower (Helianthus annuus L.) mediated by Agrobacterium rhizogenes. Zuo wu xue bao. 2006, 32 (5), 743-748.
93. Tishchenko O. M., Komisarenko A. G., Mykhalska S. I., Sergeeva L. E., Adamen-ko N. I., Morgun B. V., Kochetov A. V. Agrobacterium-mediated transformation of sunflower (Helianthus annuus L.) in vitro and in planta using Lba4404 strain harboring binary vector pBi2E with dsRNA-suppressor of proline dehydrogenase gene. Cytol. Gen. 2014, 48 (4), 218-226. doi:10.3103/ S0095452714040094.
94. Masahiro Nishihara, Takashi Nakatsuka. Genetic Engineering of Novel Flower Colors in Floricultural Plants: Recent Advances via Transgenic Approaches. Protocols for In Vitro Propagation of Ornamental Plants Methods in Molecular Biology. 2010, N 589, P. 325-347.
95. Mubarac M. Genetic Transformation in Stevia rebaudiana, International Conference On Biotechnology Applications In Agriculture, Benha University (ICBAA), Moshtohor and Hurghada, 8-12, April 2014, Egypt, 19-26.
96. Hsin-Mei Wang, Kin-Ying To. Agrobacte-rium-mediated transformation in the high-value medicinal plant Echinacea purpurea. Plant Sci. 2004, 166 (4), 1087-1096. doi: 10.1016/j.plantsci.2003.12.035.
97. Wang B., Zhang G., Zhu L., Chen L., Zhang Y. Genetic transformation of Echinacea purpurea with Agrobacterium rhizogenes and bioactive ingredient analysis in transformed cultures. Coll. Surf. B. Biointer. 2006, 53 (1), 101-104. PMID: 16982176.
Agrobacterium- 1 COMPOSITAE. I. I » 1
. .
1 i ii HAH ,
i ai ia i ax i ixix ixi, i aai Compositae Agrobacterium tumefaciens A. rhizogenes, i, ix (Cichorium intybus, Lactuca sativa), ix (Helianthus annuus), ax (Gerbera hybrida), iax (Bidens pilosa, Artemisia annua, Artemisia vulgaris, Calendula officinalis, Withania somnifera .). ii Compositae, , CTrn^x x, ax, ia, ( ii, i aix x i, i i ). Ha ixi ax» i Compositae, Cynara cardunculus, Arnica montana, Cichorium intybus, Artemisia annua.
i : Compositae, Agrobacterium tumerfaciens, Agrobacterium rhizogenes, » .
Agrobacterium- COMPOSITAE. I. »
. .
, Compositae Agrobacterium tumefaciens A. rhizogenes, , (Cichorium intybus, Lactuca sativa), (Helianthus annuus), (Gerbera hybrida), (Bidens pilosa, Artemisia annua, Artemisia vulgaris, Calendula officinalis, Withania somnifera .) Compositae, , , , ( , , ). » Compositae, Cynara cardunculus, Arnica montana, Cichorium intybus, Artemisia annua.
: Compositae, Agrobacterium tumerfaciens, Agrobacterium rhizogenes, » .