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Sunflower tissue culture.

Introduction

Sunflower (Helianthus annuus L) is the third important major edible oil seed crop in the world after soybean and groundnut. The significant developments that took place in varietal front and the remarkable ability of the crop to adjust and grow successfully in different agro climatic conditions expanded sunflower production to all the continents. Consequently the sunflower area has crossed 20 million hectares, producing around 30 million tonnes of seed annually (Table 1).

Of the different biotic stress factors, which are the bottleneck for increasing the productivity of sunflower, susceptibility to fungal diseases is a major limiting factor for increasing sunflower production in world. Of the four major diseases (viz., Alternaria blight, rust, downy mildew and root / collar rot)of sunflower, Alternaria blight caused by Alternaria helianthi (Hansf.). All open pollinated varieties in cultivation now are susceptible to Alternaria leaf spot, rust and downy mildew indicating the vulnerability of sunflower production in the world. As all the currently grown varieties and hybrids are susceptible to Alternaria leaf spot, which is widespread in the world, it is necessary to identify durable resistance and this has to be a major thrust area in resistant breeding programmes. As a first step in this direction, efforts are needed to identify the resistant sources. The germplasm screening trials to date have indicated lack of genetic resistance to this disease in cultivated sunflower. Even all other wild diploid species are susceptible. Only in some perennial tetraploid and hexaploid species particularly in H. tuberosus there are indications of existence of some resistance [28]. As the disease can cause severe leaf, stem and head spots resulting in pre-mature death and defoliation leading to yield losses of upto 80 percent under favourable environmental conditions, efficient strategies for the control of this disease are necessary.

Among options open to plant breeders to widen the genetic base, are exploitation of alien variation and somaclonal variation. It is highly pertinent to devise and adopt new and conventional means to complement the traditional methods of breeding for disease resistance [115] especially when there are no resistant sources available. The advances made in the area of plant cell and tissue culture and the eventual regeneration of entire plants from them have established this technology as a powerful tool for use in crop improvement programmes. The immense possibilities offered by the application of the technique of tissue culture for genetic upgrading of economically important plants have been emphasized by several workers [27,38,2]. By using tissue culture techniques disease resistant plants have been successfully produced [85,119,110,104,59,105,3]. Therefore, cell and tissue culture will surely complement the conventional practices and will furnish a valuable tool for agricultural research for obtaining disease resistant plants.

Information on the cultural conditions favoring callus induction and subsequent regeneration of plantlets is a' pre-requisite for the utilization of this technique. In fact the success of many in vitro techniques in higher plants depends on the success of plant regeneration [79]. The application of some techniques like selection for salinity tolerance, drought and disease resistant plants, in-vitro mutagenesis and somatic hybridization is limited in many of the crop species because of their highly recalcitrant nature and difficulties to regenerate plants especially when they are subjected to stress conditions in vitro.

Origin, distribution and importance of sunflower:

The genus Helianthus (Asteraceae) includes about 100 species, the majority of which are native to North America. The genus provides two food plants, H. annuus, the sunflower, and H. tuberosus, the Jerusalem artichoke. Several varieties of H. annuus, as well as other species of the genus, are sometimes cultivated as ornamentals. The common sunflower (H. annuus L.) is cultivated in every continent and is one of the four major annual crops grown for edible oil.

The sunflower oil is used for human consumption for its high percentage of polyunsaturated fatty acids and as a raw material for oleochemistry. It can also be used as substitute for mineral oil in various applications such as a fuel, lubricant, or an oil for hydraulic systems [71].

The Genus Helianthus, is named from Greek where 'helios' means 'Sun', 'anthus' means 'flower'. Helianthus annuus was so named by C. Linnaeus, to, when the only sunflower known, lived in a single season (annual). Sunflower probably originated in South-West United States and its seed was very early used for food by Indians. The cultivated sunflower is native of America. It was taken to Spain from Central America before "grown by Indians for food and in New England for hair oil in 1615.

Botanical description of sunflower:

Sunflower (Helianthus annuus L.) is a member of thistle family Asteraceae. Sunflower plants are herbaceous, annuals that grow from 4' to 20" (1-6m) tall, usually with only a single, hair covered stem that may be more than an inch in diameter. Leaves are as long as 12 inches (30 cm) and are borne on Petioles arranged alternately on the stem. Sunflower has a typical head type of Inflorescence. The ray florets are located around the margin of the head, 'are sterile. The corollas of the flower are the petals of the sunflower. The disc florets (flowers) located in the central part of the head are fertile but incomplete. The ovary of the disc flower develops into an achene, the same type of fruit as that of safflower.

The sunflower has peculiar response to light. In the morning the head is facing eastward, towards the rising sun; during the day, the head (or the stem tissue surrounding/supporting it) turns so that the head follows the path of the sun. At sunset, the head is facing west. During the night the head rotates towards the east and the cycle is repeated. The heliotropic response of the sunflower anticipates the appearance of the sun. Sunflower leaves display the same type of heliotropic response, and infact, they may trigger it. If the leaves are removed, the head no longer follows the sun. The plant ceases its heliotropic behaviour at, or shortly after anthesis. As a result, as much as 90% of the heads are facing east at maturity. This phenomenon simplifies harvesting. By planting sunflower in North-South rows, the vast majority of the mature heads, which end up facing east, may be hand picked from tucks moving along one side of the crop row.

Plant tissue culture:

Plant tissue culture techniques have become a powerful tool for studying basic and applied problems in plant biology. Tissue culture technology offers a great potential in addition to the traditional methods of plant breeding and improvement particularly in the area of genotype evolution with chemical tolerance and in clonal propagation. Scientists all over the world wish to apply in vitro methods for the management and breeding of economically important plants i.e. those involved in modification and improvement of plants for the production of food, fibre and fuel. With the increase in world population, the continued loss of prime agriculture land for housing and industry, necessitates the use of more marginal land for agriculture and new approaches are needed to the increasing problems such as diseases and salinity. Tissue culture technology offers one such possibility. When hlaberlandt, (1902) attempted the first plant tissue culture, his intention was to develop a more versatile tool to explore morphogenesis and to demonstrate totipotentiality of plant cells. But recent advances in the techniques and applications of plant cell culture and plant molecular biology have created unprecedented opportunities for the genetic manipulation of plants. Thus, the primary goal of plant tissue culture research is crop improvement.

The establishment of viable plant tissue culture by White, [5]; Gautheret, [82] and Nobecourt, [45] about 65 years ago marked the beginning of this rapidly developing area of basic and applied research. The discovery of role of phytohormones such as Kinetin by Skoog [10], and demonstration of totipotency of plant cells by Steward, et al., [12], auxins by Thimman & Leopold, [18], clonal propagation of orchids by Morel [55], embryo culture by Maheshwari, [62] haploids by Guha & Maheshwari, [73] protoplast isolation and fusion by Cocking [98] and Takebe, et al, [15] are the other landmarks in the development of this field.

Plant cell and tissue culture has opened a number of possibilities and much literature has accumulated with new investigations over the last three decades. The application of tissue culture techniques and the benefits derived out of it, such as production of quality plants in large quantities by clonal propagation, establishment of hybrid plants and stress resistant plants has improved the trade and industry.

Organ formation in tissue culture was observed with tobacco tissues [6] Carrot explants ^Nobecourt, [46], Ulmus cambium cultures and Witloof tissue [83]. The meristem culture work initiated by Ball, [116] was applied by Morej to. Orchids. Now this new method of vegetative propagation is employed intensively in Horticulture and nursery industry for the rapid clonal propagation of plants.

The present and future utility of plant tissue culture is closely linked to success in regenerating whole plants from cultured plant materials, but regeneration is possible in some but not all species. For successful induction of callus and subsequent regeneration of plants, proper nutrient media have to be developed and standardized. The basic nutritional requirements of cultured plant cells are very similar to those normally utilized by whole plants. To meet particular requirements for cell tissue and organs, several media were developed [39,60,99,81]. These growth media supported the growth and morphogenesis in wide varieties of plants.

Sunflower tissue culture:

The application of biotechnological methods for improving the characteristics of sunflower is limited mainly by the difficulty of regenerating plants in a reproducible and' efficient way. Sunflower regenerability by organogenesis is highly variable and depends upon genotype, specific media components and the nature of the explant. Over the last few years, a variety of techniques for regeneration by organogenesis [34,7,90,97,109,120,121] or somatic embryogenesis [93,80,90,31,36,69,44] have been described in this species. Plant regeneration parameters have been shown to be under quantitative genetic control in sunflower [25,26,101,121]). A summarised review of sunflower tissue culture is presented here decade wise.

Most reports of Helianthus annuus (L.) tissue culture prior to 1970 were concerned about crown gall cells. By the early 1950's non tumorous callus from Helianthus annuus had been induced by growing stem or hypocotyl segments on simple media with auxin [64,76,75]. The first attempts to establish sunflower callus that would differentiate into whole plants were made in 1974. Rogers et al, [20] successfully established callus from a CMS sunflower line, but only roots were differentiated from this callus. The first report of successful plant regeneration from sunflower callus was by Sadhu [24]. By growing stem pith on a modified White's medium with 1 mg/l IAA, callus was induced. After 10 weeks, one piece of callus differentiated into several plantlets.

Georgieva--Todorova et al. [86] reported regeneration from callus of the sterile hybrid H.annuus X H.decapetalus. Greco et al., [72] showed that callus capable of regeneration can be obtained from every part of the seedlings (except roots). Paterson [42] reported shoot tip culture, flowering and development of adventitious and multiple shoots in sunflower.

Paterson & Everett [41] reported embryogenesis of sunflower inbreds from 12 day old seedling hypocotyl explants on a modified MS medium (supplemented with 6.9 g/l total KN03, 40 mg/l Adenine sulphate, 500 mg/l casamino acids, 1.0 mg/l BA, 1.0 mg/l NAA and 0.1 mg/l GA3). Bohorova, et al., [112] isolated and cultured protoplasts of wild and cultivated sunflower and obtained roots and meristematic regions from protoplast derived callus. Lupi, et al., [61] established a technique for plant regeneration from shoot apex derived calli. Cavallini & Lupi [108] studied cytological condition of In vitro shoot apical meristem derived calli and regenerated shoots. Finer [93] reported direct somatic embryogenesis and plant regeneration from immature embryos of hybrid sunflower on high sucrose containing medium. Power [34] reported organogenesis from mature and immature zygotic embryos of sunflower inbreds and hybrids. Freyssinet & Freyssinet [80] produced fertile plants from hypocotyl of immature embryos from six inbred lines. McCann [53] regenerated plantlets from immature embryo derived callus from seven commercial inbred cultivars. Witrzens, et al., [7] described a method for the culture and regeneration of plants from immature embryo derived callus of sunflower hybrids. Anne-Laure Moyne, et al. (1988).

initiated callus and embryoids from hypocoyt! protoplasts of sunflower and paved the way for using protoplasts as an ideal system for sunflower regeneration in vitro. Nataraja and Ganapathi [43] obtained plant regeneration in vitro from cotyledons of sunflower Cv. Morden. Espinasse, et al., [91] studied the effect of growth regulator concentration and explant size on shoot organogenesis from callus derived from zygotic embryos of 5 sunflower genotypes. Espinasse & Lay [90] obtained shoot regeneration of callus derived from globular to torpedo embryos from 59 sunflower genotypes. Dupuis, et al, [87] studied the effect of plant donor tissue and isolation procedure effect on early embryoid formation from protoplasts of sunflower. Prado & Berville [36] reported induction of somatic embryos and their development by liquid culture of hypocotyl derived calli of 2 sunflower genotypes. Pelissier, et al, [31] produced isolated somatic embryos from hypocotyl thin cell layers of sunflower.

Knittel, et al., [47] obtained high frequency plant regeneration from mature sunflower cotyledons. Chraibi, et al, [96] reported stimulation of shoot regeneration from cotyledons of sunflower by ethylene inhibitors, silver and cobalt. Pugliesi, et al, [37] showed plant regeneration and genetic variability in tissue cultures of sunflower cotyledons. Burrus, et al, [106] regenerated fertile plants from hypocotyl protoplasts of sunflower. Jeannin & Hahne [69] studied donor plant growth conditions and regeneration of fertile plants from immature embryo derived somatic embryos of sunflower. Fischer, et ai, (1992) regenerated fertile plants derived from hypocotyl & cotyledon derived protoplasts of sunflower. Chraibi, et al., [97] reported genotype independent system of regeneration from cotyledons of sunflower and studied the role of ethylene in regeneration. Polger & Krasnyanski [32] described methods of plant regeneration from cell suspension and protoplast culture of H.maximiliani. Punia & Bohorova [30] obtained plant regeneration from callus of six wild species of sunflower using stem, leaf, bud and cotyledons, Ceriani, et al., [109] reported regeneration from cotyledons of 10 genotypes in sunflower and transformation with Agrobacterium. Krasnyanski & Menczel [50] regenerated plantlets via somatic embryos derived from hypocotyl protoplasts of sunflower. Pugliesi, et al., [29] described a method of organogenesis and embryogenesis from cotyledons and anthers of H. tuberosus and interspecific hybrid H. tuberosus X H. annuus. Thengane, et al., [17] cultured anthers of 4 genotypes and regenerated plantlets from embryos of only one genotype. They studied influence of genotype and culture conditions on embryo induction and plant regeneration.

Keller, et al., [66] studied ultrastructure of sunflower mesophyll protoplast derived callus and reported differences in their regenerative potential. Kiranmai & Prathibha Devi [67,68] studied callusing and organogenesis from cotyledons of normal and mutated sunflower. Zhong, et al., [9] reported formation of doubled hapioid sunflower plants by androgenesis using in vitro anther culture which later turned out to be derived from diploid callus of anther wall / connective tissue rather than from anthers. Coumans & Zhong [100] studied the formation of doubled hapioid sunflower plants by androgenesis using in vitro isolated microspore culture and obtained microcallus from microspores after eliminating the associated somatic tissue by a two step purification. Addition of Aminocyclopropane caboxylic acid (ACC) induced the division of microspore derived microcallus.

Krasnyanski and Menczel [51] reported production of sunflower somatic hybrid plants (H. annuus X H. giganteus) by protoplast fusion using polyethylene glycol (PEG). Jeannin, et al, [70] studied the role of sugar on somatic embryogenesis and organogenesis induced on immature zygotic embryos of sunflower cultivated in vitro. Laparra, et al., [56] obtained plant regeneration from different explants in Helianthus smithii, a wild species using leaf, rhizome and stem. Mita, er ai, [54] reported presence of secreted heat shock proteins in sunflower suspension cell cultures. Fambrini, et al., [92] reported acquisition of high embryogenic potential in regenerated plants of H. annuus X H. tuberosus, a tetraploid interspecific hybrid. Keller, et al, [65] studied the influence of electrical treatment and cell fusion on cell proliferation capacity of sunflower protoplasts in very low-density culture. Barthou et al., [117] studied the effect of atmospheric pressure on sunflower protoplast division. Fiore, et al., [94] reported high frequency plant regeneration in sunflower from cotyledons via somatic embryogenesis of several genotypes. Henn et al., [78] regenerated fertile interspecific hybrids from protoplast fusions between H. annuus and wild HeJianthus species. Henn, et ai, [77] regenerated fertile plants from H. nuttallii and H. giganteus mesophyll protoplasts. Charhere & Hahne (1998) studied induction of embryogenesis versus caulogenesis on in vitro cultured sunflower immature zygotic embryos with respect to role of plant growth regulators, sugar and auxin polar transport inhibitors. konov, et al [49] reported formation of epiphyllous buds' in sunflower and studied its induction in vitro and cellular origin. Berrios, et al., [121] studied the influence of genotype and gelling agents on in vitro regeneration by organogenesis in 14 recombinant inbred lines and their parents of sunflower. Berrios, et al., [120] evaluated the genetic variability for regenerability by organogenesis in 93 recombinant inbred lines, of sunflower. Potdar, et ai, [33] repored genotype dependent somatic embryogenesis in sunflower. Pandurang [40] reported selection and regeneration of salinity tolerant cell lines in Sunflower (H. annuus L) through cell and tissue culture. Sujatha & Prabhakaran [40] reported in vitro capitulum induction on shoot cultures of cultivated and wild species of sunflower. Weber, et al., [4] reported high regeneration potential in vitro from apical meristems of sunflower lines derived from interspecific hybridization. Sujatha and Prabhakaran [14] obtained high frequency embryogenesis from immature zygotic embryos of sunflower and studied the influence of various factors like genotype, embryo size, sucrose concentration carbohydrate source, agar strength, basal media, photoperiod and temperature effect on embryogenesis. reported micropropagation of H. maximiiiani by shoot apex culture. Yordanov, et al., [8] obtained plant regeneration from interspecific hybrids and backcross progeny of Helianthus eggertii X H. annuus. Dhaka and Kothari [102] reported improved bud elonngation and in vitro plant regeneration efficiency in sunflower using Phenyl acetic acid (PAA).

Selection for in vitro disease resistance in plants:

One of the earliest examples of selection for disease resistance exploited the structural similarities between an amino-acid analogue methionine sulphoximine and a toxin produced by Pseudomonas tabaci. Carlson [107] was the first to produce tobacco plants from the callus cultures which were resistant to Methionine sulphoximine, an analogue of tobaxin, produced by P. tabaci and enhanced resistance to the pathogen was observed. Initially, Bajaj and Saettler [114] had studied the effect of Halotoxin containing filtrates of Pseudomonas phaseoiicola on the growth of bean callus tissue. Krishnamurthi and Thaskal [52] reported Fiji disease resistant sugarcane subclones {Saccharum officinarum Var. Pinder) from tissue culture. successfully selected soybean (Glycine) plants from the suspension cultured soybean cells to the elicitor isolated from the fungal pathogen Phytophtora megasperma var. Sojae. Gengenbach, et al. [85] reported maize plants resistant to Helminthosporium maydis. Sacristan & Hoffman [22] studied direct infection of embryogenic tissue cultures of haploid Brassica napus with resting spores of Plasmodiophora brassicae. Earle, et al., [88] studied the effect of Helminthosporium phytotoxins on cereal leaf protoplasts. Chaleff [100] isolated sugarcane plants resistant to eyespot disease from callus and suspension culture, subjected to toxin of Helminthosporium sacchari, about 15-20% regenerated plants proved resistant. Behnke [118,119] selected potato plants resistant to un-fractioned liquid fungal growth medium containing exotoxins of Phyophthora infestans. Bajaj, et al., [115] studied differentiated tolerance of tissue cultures of pearl millet to Ergot extract. Larkin and Scowcroft [58] reported regeneration of sugarcane clones resistant to Helminthosporium sacchari from single cell population of susceptible clone treated with Heiminthosporoside. Gengenbach, et al, [84] studied mitochondrial DNA variation in maize plants regenerated during tissue culture selection. Sacristan [23] selected Phoma lingum resistant plants of Brasslca napus from cell and embryonic cultures. Thanutong, et al. [16] reported resistant tobacco plants from protoplast derived callus selected for their resistance to Pseudomonas & Alternaria toxins. Solodkaya & Mezentsev [11] regenerated clover plants from a cell suspension cultivated in a medium containing the culture liquid of Sclerotinia trifollium Erikss. fungus. Larkin and Scowcroft [57] studied somaclonal variation and eye spot toxin tolerance in sugarcane. Bhagyalakshmi, et al, [122] obtained downy mildew resistant pearl millet plants from infected tissues cultured in vitro Brettel, et al, [117] reported regeneration from callus treated with toxic substances produced by Leptosphaeria nodonum Mutler, which cause Glume blotch disease of wheat. Maribona, et al, [48] isolated sugarcane callus culture resistant to Helminthosporium sacchari toxin. Scala, et al., [21] analysed the Tomato--Fusarium oxysporum system for the selection of disease resistance. The effect of selective toxin of Helminthosporium victoriae on oat tissues and protoplasts was studied by Briggs, et al [104] and they were able to select Helminthosporium resistant plants from oat callus. Similarly, Rines & Luke [35] successfully selected oat plants insensitive to Helminthosporium victoriae toxin. The toxic effect of Fusaric acid on mesophyll cells of Asparagus officinalis in cell cultures was studied by Julian in [63]. Ling, et al., [59] reported rice plants resistant to Helminthosporium oryzae through tissue culture.

Refrences

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Mohammad Sharrif Moghaddasi Islamic Azad University, Saveh branch, Iran

Mohammad Sharrif Moghaddasi: Sunflower Tissue Culture

Corresponding Author

Mohammad Sharrif Moghaddasi, Islamic Azad University, Saveh branch, Iran

E-mail: Memo1340@yahoo.com
Table 1: Sunflower seed Production, Harvested area
and Yield Production in countries(2009 FAO).

Country Harvested area (Ha) Production Yield (Kg/ha)

Russian federation 5.597.900 6.454.320 11.529
Ukraina 4.193.000 3.360.600 15.169
Argentina 1.820.030 2.483.437 13.645
Bulgaria 683.711 1.317.979 19.276
China 97.000 1.650.000 17.010
France 724.800 1.703.900 23.508
Hungary 535.090 1.256.185 23.476
U.S.A 790.562 1.377.130 17.419
India 1.800.000 1.440.000 5.800
Croatia 23.366 82.098 30.000
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Title Annotation:Original Article
Author:Moghaddasi, Mohammad Sharrif
Publication:Advances in Environmental Biology
Article Type:Report
Geographic Code:7IRAN
Date:Mar 1, 2011
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