Nitrogen Fixation as a Stress-Avoidance Strategy Among Actinorhizal (Non-Legume) Trees and Shrubs

Summary

Actinorhizal species of trees and shrubs are a diverse group of plants that may survive in poor soils by virtue of their associations with the actinomycete Frankia. These species include several important woody plants that are well suited for horticultural use in temperate climates. The symbiosis between Frankia and actinorhizal species shows some similarity to symbioses between rhizobia and woody legumes, and a common ancestor has been proposed for the predisposition to root-nodule symbiosis. Despite their probable common origin, important differences exist between actinorhizal and leguminous symbioses; characteristics of the microsymbiont, nodule architecture, and mechanisms controlling oxygen relations of the nodule are among the ways the two systems differ. If nitrogen fixation is sustained under unfavorable conditions, woody plants that associate with nitrogen-fixing organisms may show enhanced tolerance of environmental stress; species of plants capable of nitrogen-fixing symbioses are known to have comparatively strong resistance to invasion by pathogens. Expansion of the capacity to form nitrogen-fixing symbioses to novel species is a goal of those concerned with the economic and ecological impact of chemical fertilizers. Small in-roads have been made in this regard, but much remains to be discovered about introducing nitrogen fixation to additional species. Herein we review biological aspects of actinorhizal symbioses; consider the horticultural potential of temperate, woody species that form these symbioses; and discuss how nitrogen-fixing symbiosis may impact the stress resistance and use of actinorhizal species as horticultural crops.     

Engineering for Drought Tolerance in Horticultural and Ornamental Plants

Abstract

Drought is one of the major factors limiting plant growth and productivity. Plant adaptation to drought is dependent on molecular networks for drought perception, signal transduction, expression of a subset of genes and production of metabolites that protect and maintain the structure of cellular components. In general, the drought response pathways can be classified into two categories: one is dependent on the stress hormone abscisic acid (ABA) and the other is ABA-independent. Many genes in these pathways have been identified, thereby providing guidance in choosing genes for engineering of drought tolerance. The review highlights the genes that mediate drought response and tolerance, and discusses lessons learned from engineering for drought tolerance in model plants, such as Arabidopsis, rice and tobacco. Because success of drought tolerance engineering is dependent on not only protein coding regions but also appropriate promoters, this article also reviews the promoters that are crucial for successful engineering of stress tolerance.     

Ornamental Plant Transformation

Abstract

Ornamental plant transformation has advanced considerably in the last decade. Now over 40 genera have been reported to be transformed. The primary methods of creating transgenic ornamental species have been Agrobacterium tumefaciens-medmtedtransformation and microprojectile bombardment. The vast majority of reports indicate the use of Agrobacteriummedmtedtransformation employing binary vectors and virhelper plasmids or supervirulence genes. Many reports are of transformation with the uidA reporter gene driven by the 35S cauliflower mosaic virus promoter, but recent efforts are now focusing on trait genes including disease resistance, flower color, flower longevity, floral scent and plant habit. Greater use of tissue specific and inducible promoters promises to enhance the functionality and usefulness of introduced trait genes. While technical challenges for production of transgenic ornamental plants still exist, the greatest challenges to realizing the potential benefits of transgenic ornamental plants are questionable public acceptance of transgenic plants and the prohibitive costs of generating environmental impact data needed to gain regulatory clearance.     

农杆菌介导的转ICE1基因提高水稻的耐寒性

利用农杆菌介导的转基因技术,成功地将通过RT PCR克隆的拟南芥ICE1基因导入垦鉴稻10号中,经PCR和Southern检测确认目的基因已整合到水稻基因组中。潮霉素抗性测定结果表明,与未转基因水稻相比,T1代表现出对潮霉素较高的抗性和孟德尔式的单位点遗传。抗寒能力检测结果表明,在同等低温胁迫条件下T1代转基因株系的死亡率明显低于未转基因对照。脯氨酸含量增幅明显高于未转基因对照。上述结果表明,转ICE1水稻提高了抗寒能力。     

Effect of Dwarfing Induced by Uniconazole-P on Snow Tolerance of the Faba Bean (Vicia faba L.)

Abstract

To clarify the effect of artificial dwarfing induced by uniconazole-P, a gibberellin biosynthesis inhibitor, on the snow tolerance of faba bean, we examined the snow damage, non-structural carbohydrate (NSC) concentration, and resistance to snow molds of the seedlings treated with uniconazole-P. Artificial dwarfing markedly decreased snow damage caused by continuous snow cover for 62 days. NSC concentration, which affects physiological snow tolerance and snow mold resistance, was lower in dwarfed plants than in non-dwarfed plants. However, there was no difference in the rate of decrease of NSC concentration between dwarfed and non-dwarfed plants under cold and dark conditions simulating snow cover. In inoculation tests, the lesions of Pythium iwayamai and Sclerotinia trifoliorum on the leaves detached from dwarfed plants were shorter than those on the leaves from non-dwarfed plants. In artificially dwarfed plants, the leaves were thicker and darker in color, and mesophyll cells were larger and denser than those in the non-dwarfed plants. Our findings suggest that the artificial dwarfing caused by uniconazole-P increased the resistance of the leaves to snow molds, thereby increasing snow tolerance, without increasing NSC concentration. The microstructural changes that occur with dwarfing may be a factor in snow tolerance.     

Molecular Approaches to Improve Salt Resistance in Crops: Facts and Perspectives

SUMMARY

Because of the expansion of agriculture into marginal environments, enhancement of crop resistance to soil salinity is becoming a frequent objective for breeders. The tools offered by molecular biology to transfer a single or a few genes provide a major hope to reduce the negative impact of broad gene transfer that takes place in wide-cross hybridizations. Due to the presence of osmotic and toxic components in the growth response of plants to salt stress, any attempt to improve plant performance in saline environments should ensure the maintenance of an adequate flux of water into plant tissues, and also avoid the build up of ions into the cell compartments where they can exert toxic effects. Besides, reduction of injury effects due to salinity on plant tissues is a highly desirable objective. Transgenic plants overexpressing ion transporters able to exclude Na⁺ into vacuoles, the enzymes required for the biosynthesis of several osmocompatible, organic solutes, or the enzymes participating in detoxification pathways, have been obtained. Some of these transgenic plants display an enhanced growth relative to their wild type parents in saline environments, although the way in which this resistance is achieved remains essentially unknown. A fourth and promising way to engineer salt resistance in plants is the attempt to manipulate gene regulatory pathways. The extent to which these experiences, mainly with model plants, could be extrapolated to crop plants growing in the field is discussed. It is proposed that a combination of different molecular approaches could be helpful to achieve enhanced salt resistance in crop plants.     

TILLING: an alternative path for crop improvement

Rising global temperatures, changing precipitation patterns, spread of insects and diseases, and slow development of crop varieties acclimatized to new environments have produced a food crisis situation in some developing countries. Although transgenic technologies came to the rescue, they faced numerous hurdles on the journey to reach the farm soil, which has necessitated innovation of out-of-the-box technologies for crop improvement. One such non-transgenic method, which exploits the ever-increasing sequence data and combines it with traditional and modern methods to induce and extract mutants, is Targeting Induced Local Lesions IN Genomes (TILLING). With the success of TILLING in monocots, dicots, autopolyploids, allopolyploids, and triploids, we expect this technique to help develop many mutant crop varieties in the next few decades. We trust this review will be useful for researchers engaged in crop improvement around the globe.     

Freeze tolerance and physiological changes during cold acclimation of giant reed [Arundo donax (L.)]

Arundo donax L. (Poaceae), giant reed, is a rhizomatous shrubby grass that is cultivated in subtropical and warm temperate regions for a multitude of uses. Recently, it has been identified as a leading sustainable, non‐food crop for lignocellulosic biofuels due to its low ecological and agronomic demands. Lack of cold hardiness may limit its diffusion into colder areas of the transition zone. The objectives of this study were to (i) quantify freeze tolerance (LT₅₀) of cold‐acclimated and non‐acclimated Arundo donax L. plants using reproducible, controlled environment conditions, (ii) determine the effect of prolonged exposure to freeze stress on tolerance by keeping plants at a constant, sublethal temperature and (iii) study the relationship of non‐structural carbohydrates (total soluble sugars, glucose, fructose and sucrose) and proline accumulation with cold hardiness. In vitro‐propagated plants of the Honduran and Hungarian ecotypes of Arundo donax L. were chosen for this study. Cold acclimation treatment was imposed for 1 week using a controlled environment chamber set at 10°C and with a 12‐h photoperiod of 200 μmol m^?2 s^?1 photosynthetically active radiation. Freeze tolerance ranged from ?12·8°C (Honduran) to ?16·4°C (Hungarian ecotype). In all the organs analysed, total soluble sugars significantly increased during cold acclimation, with concentrations between 1·8‐ and 4·7‐fold higher than in non‐acclimated plants. The higher concentrations of sugars and proline in cold‐acclimated plants were positively associated with enhanced giant reed freeze tolerance (2·9°C lower). Our results confirm that during cold acclimation, metabolic changes related to increased freezing tolerance occur in giant reed.     

Inheritance of Cold Hardiness and Dehydrin Genes in Diploid Mapping Populations of Blueberry

Summary

Studies of herbaceous plants suggest that cold hardiness is a complex, quantitatively inherited trait. Although development of cold hardiness is an integral part of the life cycle of woody perennial plants, studies on the genetic control of cold hardiness in woody perennials are scarce. A better understanding of the genetic control of cold hardiness would be valuable for developing more effective strategies to increase cold hardiness and, hence, climatic adaptation of woody perennial crops. In blueberry, three major dehydrins of 65, 60, and 14 kDa have been found to increase with cold acclimation and decrease with deac-climation. A comparison of these dehydrin levels among various blueberry cultivars and selections has revealed their level of accumulation to be closely associated with cold hardiness level. Efforts are underway to isolate and map the dehydrin genes of blueberry utilizing blueberry populations that segregate for cold hardiness in order to determine if the dehydrin genes map to or co-segregate with QTLs controlling cold hardiness. Progress has been made toward this goal. Cold hardiness levels were determined for a portion of the blueberry mapping populations (derived from testcrosses of Vaccinium darrowi Camp X V. caesariense Mackenz. F₁s to another V. darrowi and another V. caesariense) using a laboratory controlled freeze-thaw regime, followed by visual assessment of injury to floral buds. As expected, the V. darrowi and V. caesariense parents were found to differ significantly in terms of cold hardiness levels (LT₅₀s of -13°C and -20°C, respectively). Mean cold hardiness level of F₁s (LT₅₀ of -14.7°C) was skewed toward the V. darrowi parents suggesting that cold hardiness is a partially recessive trait. The sequence of a 2.0 kb cDNA clone, which encodes the 60 kDa blueberry dehydrin, was used to map a dehydrin-related gene to current linkage group 12 of the V. caesariense testcross population. A preliminary comparison of the segregation pattern of the dehydrin-related gene to that of the cold hardiness trait suggests that the marker does not segregate with cold hardiness.     

Bacillus thuringiensis cry1Ab gene confers resistance to potato againstHelicoverpa armigera (Hubner)

Summary Helicoverpa armigera is one of the important insect pests adversely affecting the yield of potatoes in India. A synthetic gene encoding the insecticidal crystal protein (Cry1Ab) ofBacillus thuringiensis (Bt) has been introduced into five genotypes of potato usingAgrobacterium tumefaciens. Southern analysis of DNA from transgenic plants confirmed the integration and copy number of the transgene. Double-antibody quantitative sandwich ELISA analysis demonstrated high levels of Cry1Ab protein expression in transgenic plants. Insect bioassays on the leaves of transgenic plants showed considerable protection against the larvae ofH. armigera in terms of leaf area consumed and larval weight reduction.     

Improvement of Horticultural and Ornamental Crops Through Transgenic Manipulation of the Phytochrome Family of Plant Photoreceptors

Abstract

Consumer demands drive continuous developments in the production of horticultural and ornamental crops. In addition to improvements in quality and nutritional content, crop producers must supply an increased variety of products to a year-round market. The availability of many horticultural and ornamental crop products is dependent on the timing of reproductive development. The time at which many plants initiate sexual or vegetative reproduction is governed by a number of interacting environmental factors such as daylength, light quality and temperature. Artificial manipulation of the growing environment is therefore frequently used to ensure production meets retailers' marketing programs, a strategy which can often result in high energy costs. An alternative approach involves the manipulation of genes encoding proteins responsible for perceiving and transducing environmental stimuli, in particular, the genes encoding the phytochrome family of plant photoreceptors. Alterations in the expression of genes encoding phytochromes can modulate not only the timing of reproductive development, but also plant architecture. Such approaches can therefore be used to modulate a variety of phenotypic traits such as height, lateral branching and harvest yield, while enabling growers to tailor crop reproduction to their marketing needs. In this review, we will discuss examples of crop improvement using transgenic manipulation of phytochrome expression, along with benefits and disadvantages of such approaches.     

The Development of Coleoptile Tillers in Relation to Seedling Vigor in Early-Maturing Varieties of Spring Type Wheat

Abstract

Increase in seedling vigor is necessary to improve the yield of early-maturing spring wheat with a limited growth period. The importance of coleoptile tillers in increasing seedling vigor has been suggested. However, most previous studies on the development of coleoptile tillers have been made with only a limited number of varieties and under controlled environmental conditions. In this study, we examined difference in the development of coleoptile tillers among early-maturing varieties of wheat under field conditions. We observed large differences among varieties in the percentage of plants with coleoptile tillers, ranging from 5.6% in Haruyutaka to 60.1% in Bandouwase. The percentage of plants with coleoptile tillers was closely correlated with the number of leaves developed on the main stem. The dry weight of the plants with coleoptile tillers was heavier than that without coleoptile tillers (by 34.8% in Fukuwase komugi). However, in some varieties, the dry weight of the plants with coleoptile tillers was similar to that of plants without coleoptile tillers. The increase in dry weight per plant seemed to be achieved by the production of either a few heavy tillers or many light tillers.     

转铁蛋白基因增强水稻对氧化胁迫与稻瘟病菌的抗性

对转豌豆铁蛋白（pea ferritin,Fer）基因水稻T1代的53个株系进行PCR检测,52个株系能扩增出阳性PCR产物。通过测定光合作用过程中最大光化学通量（Fv/Fm值）分析了由百草枯处理引起的T1代水稻叶片的氧化损害。与未转基因水稻相比,转Fer基因水稻的叶片对氧化胁迫的耐受能力有不同程度的增强。百草枯处理后转基因植株叶片叶绿素含量与水处理叶片相比没有明显下降,而未转基因植株叶片叶绿素含量降低至水处理叶片的20％左右。选取9株对氧化胁迫耐受能力较强的水稻进行了Northern blot分析和子代的稻瘟病抗性测定,其中5株转基因植株Fer mRNA 积累增强。病原菌接种后T2代转基因植株的病斑数量明显少于非转基因植株。表明转Fer基因水稻对氧化胁迫和病原菌有较好的抗性。     

转异苞滨藜BADH基因玉米高世代株系苗期外源基因表达及耐盐性功能分析

以转来自耐盐植物异苞滨藜的甜菜碱醛脱氢酶基因(BADH)玉米T8代株系BZ-136及受体对照自交系郑58(耐盐)为试材,采用盆栽种植方法,分析转基因植株中外源基因的表达情况,检测耐盐相关生理指标。结果表明,转基因株系BZ-136中的BADH酶活性及甜菜碱含量显著高于受体对照,随着盐胁迫时间的延长,其表达量先增加后减少,在胁迫7 d时分别达到了最大值。转基因株系幼苗株高和干重从盐胁迫第3天开始显著高于对照,鲜重和含水量在胁迫第5天和第7天时转基因株系显著高于对照;转基因株系根总体积显著高于对照,直径和根尖数在胁迫第5天时显著高于对照;转基因株系电导率和丙二醛含量均低于对照,分别在胁迫第3天和第7天开始达显著差异,叶绿素含量高于受体对照,从第3天开始二者差异达显著水平。由于外源BADH基因的表达显著提高了基因株系的耐盐性。     

美国转基因玉米的生产概况和发展趋势

美国是世界上最大的玉米生产国, 种植面积、总产和单产均居世界首位。转基因玉米自1996年引入生产以来,使玉米生产上了一个新台阶。当前推广利用的转基因玉米品种基本上是抗虫和抗除草剂两种类型。抗虫是由在玉米体内表达Bacillus Thuringiensis(Bt)毒素结晶蛋白(cry蛋白)基因所致,除草剂的抗性(Herbicide tolerance, HT)是由外源的或改良的EPSPS基因在玉米体内表达来实现。转基因玉米的种植面积每年都稳步上升,转基因玉米的也由单一的Bt或HT转变为多基因多性状的叠加,将抗玉米螟和抗Rootworm结合,使玉米的地上部和地下部都得到了有效的保护,再加之抗除草剂基因的叠加,使得转基因品种在生产上更具竞争力。转基因玉米的抗虫和抗除草剂等优良性状带来了巨大的经济和环境效益, 并被越来越多的大众所接受。转基因玉米在短短10年间的应用和发展,还不能回答转基因作物对人类和环境的长期影响等问题。因此,对转基因作物的管理和利用仍不能掉以轻心,不能因对生物技术的热情而对人类和环境造成无意的不可挽回的破坏。     

玉米花粉管通道法转化脱水素BDN1基因

通过花粉管通道法将脱水素基因BDN1转入玉米自交系合344中,并对转化后代进行PCR和RT-PCR分子检测以及耐盐性功能鉴定,筛选耐盐性较高的转基因玉米新种质.结果表明,实验共获得88株T0代除草剂抗性植株,其中31株PCR检测呈阳性,14株RT-PCR检测呈阳性,对T4代转基因株系苗期进行300 mmol/L NaCl溶液的盐胁迫处理, 2个转基因株系耐盐性比对照提高两个级别.     

Increased Cell-Wall Mass and Resistance to Freezing and Snow Mold during Cold Acclimation of Winter Wheat under Field Conditions

Abstract

Accumulation of soluble carbohydrates plays an important role in enhancement of resistance to freezing and snow mold of plants during cold acclimation. Nevertheless, few studies have examined whether changes in cell wall properties are involved in enhancement of resistance during cold acclimation. In this study, four winter wheat cultivars were sown in a field on six different dates during August–October, and their resistance to freezing and snow mold were compared in relation to soluble carbohydrate content and cell-wall mass in leaves. Resistance to freezing and snow mold was much higher in the plants sown on 23 September than in those sown on 9 September. The percentage of cell-wall mass in leaf to total dry mass (%CW) and water-soluble carbohydrate content also increased considerably during 9–23 September. Multiple regression analyses revealed that %CW contributed significantly to freezing resistance, whereas total water-soluble carbohydrate content contributed significantly to snow mold resistance. These results suggest that increased %CW enhances freezing resistance during cold acclimation.     

The Economics of Horticultural Biotechnology

Abstract

Technological change has driven economic progress in agriculture and will continue to play a crucial role in die 21st century. The latest wave of technological change in agriculture is based on die ability to specifically modify crop genetics dirough recombinant DNA techniques. Biotech crop varieties have been adopted on a wide scale in some agronomic crops, but horticultural crops face hurdles for commercialization. Market barriers are blocking die production of biotech horticultural crops diat have already been developed. High costs for research, development and regulatory approval combined with the small acreages planted and the diversity of varieties limit die potential for profitable applications of biotechnology for many fruits and vegetables, tree fruits and nuts, and nursery and landscape crops. Like most technological changes in agriculture, die introduction of modern biotechnology has met with spirited political opposition from some quarters. Such opposition could discourage adoption of biotech products that are wanted by some consumers, can be profitable for growers and could reduce the environmental impacts of crop production.