Application of recombinant DNA technology to plant protection: molecular approaches to engineering virus resistance in crop plants

Developments in plant tissue culture, plant transformation and regeneration, and improvements in techniques to isolate and manipulate viral genes have led to the exploitation of the concept of cross protection: turning the virus onto itself and controlling it with its own genes. By introducing and expressing genes of viral origin in crop plants, scientists have engineered resistance to several plant viruses. Some of the approaches, used singly or in combination, include expression of viral-coat protein, untranslatable sense or antisense RNA, satellite RNA, virusspecific neutralizing antibody genes, plant viral replicase, protease or movement proteins and defective, interfering RNA. All of these approaches have resulted in manifestation of virus resistance to varying degrees in several commercially important crop plants. This review summarizes the recent advances in engineering virus resistance using the above approaches, and lists specific examples of their use in cultivated crop plants of economic importance.H.R. Pappu and C.L. Niblett are with the Plant Pathology Department, University of Florida. Gainesville, FL 32611-0680, USA; R.F. Lee is with the Citrus Research and Education Center, University of Florida, Lake Alfred, FL 33850, USA. Florida Agricultural Experiment Station Journal Series No. R-04558.     

Carnivorous pitcher plants: insights in an old topic

Plant insect interactions are usually recognized as a scenario where herbivorous insects feed on a host plant. However, also the opposite situation is known, where plants feed on insects. Carnivorous pitcher plants of the genus Nepenthes as well as other pitcher plants obtain many nutrients from caught insect prey. Special features of the pitcher traps’ surface are responsible for attraction and trapping insects. Once caught, the prey is digested in the fluid of the pitchers to release nutrients and make them available for the plant. Nutrients are taken up by special glands localized on the inner surface of the pitchers. These glands also secrete the hydrolyzing enzymes into the digestion fluid. Although this is known for more than 100 years, our knowledge of the pitcher fluid composition is still limited. Only in recent years some enzymes have been purified from the pitcher fluid and their corresponding genes could be identified. Among them, many pathogenesis-related proteins have been identified, most of which exhibiting hydrolytic activities. The role of these proteins as well as the role of secondary metabolites, which have been identified in the pitcher fluid, is discussed in general and in the context of further studies on carnivorous plants that might give answers to basic questions in plant biology.     

Association of morphological and biochemical features of mulberry lines with resistance to bacterial leaf spot

Xanthomonas campestris pv. mori causes bacterial leaf spot (BLS) of mulberry (Morus spp.) worldwide. Twenty mulberry lines with contrasting response to BLS showed significant (p?<?0.01) variation between resistant and susceptible groups for stomata frequency (SF), trichome density (TD), total soluble sugar (TSS) and polyphenol (PP) content, while non-significant for chlorophyll content. Heritability (broad-sense) estimates obtained from the variance component analysis were high (≥0.72) for these traits. The disease severity index values of the lines were significantly negatively correlated with TD (r?=??0.66; p?<?0.01), TSS (r?=??0.73; p?<?0.01) and PP (r?=??0.61; p?<?0.01), while positively correlated with SF (r?=?0.65; p?<?0.01). The cluster analysis was based on highly significant four variables and DSI values, and classified the lines into two major clusters. The resistant lines that come under cluster-I had high TD, TSS and PP, while those in susceptible category had high SF in cluster-II. These findings suggest that direct selection for SF, TD, TSS and PP will be effective for development of host resistance against BLS in mulberry.     

Fungal volatiles: an environmentally friendly tool to control pathogenic microorganisms in plants

Fungi are an extraordinary and immensely diverse group of microorganisms that colonize many habitats even competing with other microorganisms. Fungi have received recognition for interesting metabolic activities that have an enormous variety of biotechnological applications. Previously, volatile organic compounds produced by fungi (FVOCs) have been demonstrated to have a great capacity for use as antagonist products against plant pathogens. However, in recent years, FVOCs have been received attention as potential alternatives to the use of traditional pesticides and, therefore, as important eco-friendly biotechnological tools to control plant pathogens. Therefore, highlighting the current state of knowledge of these fascinating FVOCs, the actual detection techniques and the bioactivity against plant pathogens is essential to the discovery of new products that can be used as biopesticides.     

Community response to transgenic plant release: using mathematical theory to predict effects of transgenic plants

Predicting the potential effects of introductions of plants on the structure of plant communities has been elusive. I suggest that mathematical models of resource competition might be useful for identifying categories of plants that either are unlikely to alter community structure or that have the potential for altering community structure. Assuming that the transgenic plant will escape and establish viable populations in nontarget habitats, this theory suggests that species that have a high minimum resource requirement are unlikely to alter community structure. The theory is elaborated to evaluate the potential effects on community structure of transgenic plants with resistance to primary consumers. Results indicate that the greatest reduction in the minimum resource requirement caused by resistance will occur when consumers are consuming enough plant biomass that the plant can no longer grow. If resistance to such a consumer were incorporated into a plant, it could lower the minimum resource requirement sufficiently that a transgenic plant would be able to alter community structure substantially. Examples of introductions of exotic plants, plant pathogens, and insect herbivores are given to support the conceptual basis of the theory. Not all transgenic plants with resistance, however, have the potential to alter community structure. Resistance to primary consumers that strongly reduce the biomass producing ability of a plant will probably be able to alter community structure, whereas resistance that reduces most other types of yield loss is less likely to alter community structure. The theory should be elaborated to incorporate more-realistic assumptions, such as those regarding reproduction, dormancy, and dispersal of the transgenic plants, and provide more detailed characterization of the potential hazard of transgenic plants to plant communities.     

Silicon-augmented resistance of plants to herbivorous insects: a review

Silicon (Si) is one of the most abundant elements in the earth's crust, although its essentiality in plant growth is not clearly established. However, the importance of Si as an element that is particularly beneficial for plants under a range of abiotic and biotic stresses is now beyond doubt. This paper reviews progress in exploring the benefits at two‐ and three‐trophic levels and the underlying mechanism of Si in enhancing the resistance of host plants to herbivorous insects. Numerous studies have shown an enhanced resistance of plants to insect herbivores including folivores, borers, and phloem and xylem feeders. Silicon may act directly on insect herbivores leading to a reduction in insect performance and plant damage. Various indirect effects may also be caused, for example, by delaying herbivore establishment and thus an increased chance of exposure to natural enemies, adverse weather events or control measures that target exposed insects. A further indirect effect of Si may be to increase tolerance of plants to abiotic stresses, notably water stress, which can in turn lead to a reduction in insect numbers and plant damage. There are two mechanisms by which Si is likely to increase resistance to herbivore feeding. Increased physical resistance (constitutive), based on solid amorphous silica, has long been considered the major mechanism of Si‐mediated defences of plants, although there is recent evidence for induced physical defence. Physical resistance involves reduced digestibility and/or increased hardness and abrasiveness of plant tissues because of silica deposition, mainly as opaline phytoliths, in various tissues, including epidermal silica cells. Further, there is now evidence that soluble Si is involved in induced chemical defences to insect herbivore attack through the enhanced production of defensive enzymes or possibly the enhanced release of plant volatiles. However, only two studies have tested for the effect of Si on an insect herbivore and third trophic level effects on the herbivore's predators and parasitoids. One study showed no effect of Si on natural enemies, but the methods used were not favourable for the detection of semiochemical‐mediated effects. Work recently commenced in Australia is methodologically and conceptually more advanced and an effect of Si on the plants' ability to generate an induced response by acting at the third trophic level was observed. This paper provides the first overview of Si in insect herbivore resistance studies, and highlights novel, recent hypotheses and findings in this area of research. Finally, we make suggestions for future research efforts in the use of Si to enhance plant resistance to insect herbivores.     

Plant pleiotropic drug resistance transporters： Transport mechanism,gene expression,and function

Pleiotropic drug resistance （PDR） transporters belonging to the ABCG subfamily of ATP-binding cassette （ABC） transporters are identified only in fungi and plants. Members of this family are expressed in plants in response to various biotic and abiotic stresses and transport a diverse array of moleculesacross membranes, Although their detailed transport mechanism is largely unknown, they play important roles in detoxification processes, preventing water loss, transport of phytohormones, and secondary metabolites. This review provides insights into transport mechanisms of plant PDR transporters, their expression profiles, and multitude functions in plants.     

Plant pleiotropic drug resistance transporters:Transport mechanism,gene expression,and function

Pleiotropic drug resistance(PDR) transporters belonging to the ABCG subfamily of ATP-binding cassette(ABC)transporters are identified only in fungi and plants. Members of this family are expressed in plants in response to various biotic and abiotic stresses and transport a diverse array of molecules across membranes. Although their detailed transport mechanism is largely unknown, they play important roles in detoxification processes, preventing water loss, transport of phytohormones,and secondary metabolites. This review provides insights into transport mechanisms of plant PDR transporters, their expression profiles, and multitude functions in plants.     

植物适应铝毒胁迫的生理及分子生物学机理

铝毒是酸性土壤上限制作物生长最重要的因素,严重影响着全世界和中国大约40％和21％耕作土壤的作物生产．近几十年来,世界各国针对植物的铝毒及其耐铝机制进行了大量的研究,并取得了较大进展．文中重点综述了植物适应铝胁迫基因型差异筛选方法及其鉴定技术、植物适应铝胁迫的生理基础及分子生物学机制等方面的研究进展,简要讨论了今后的研究方向．     

Evaluation of cowpea mini core accessions for resistance to flower bud thrips Megalurothrips sjostedti Trybom (Thysanoptera: Thripidae)

The flower bud thrips, Megalurothrips sjostedti Trybom (Thysanoptera: Thripidae), is an economically important pest of cowpea in sub‐Saharan Africa. Varietal resistance is the most preferred, environmentally friendly, cost‐effective and sustainable option for controlling this pest. The objective of this study was to identify sources of resistance to M. sjostedti among mini core accessions from the largest world cowpea germplasm collection maintained at the International Institute of Tropical Agriculture (IITA). The study was conducted during the 2015 and 2016 cropping seasons where 365 accessions were screened under field conditions. Each accession was rated visually for thrips damage score, flower abortion rate, number of pods per plant and number of thrips per flower. The resistance levels observed in genotypes TVu8631, TVu16368, TVu8671 and TVu7325 were similar to that of the resistant check “Sanzisabinli” (called Sanzi) during both seasons. In addition, 56 mini core genotypes showed moderate resistance to thrips damage. High heritability values were associated with thrips damage scores at 65 days after planting (0.60), percentage of effective peduncles (0.59), flower bud abortion rate (0.59), number of pods per plant (0.51) and number of peduncles with pods (0.5). The accessions identified with good levels of resistance to flower bud thrips will be used in cowpea breeding programs to develop improved resistant varieties.     

Interspecific hybridization of plants and resistance to herbivores: hypotheses,genetics, and variable responses in a diverse herbivore community

We studied the morphology, molecular genetics, and hebivory of two species of willows (Salix sericea and S. eriocephala) and their interspecific hybrids to test four alternative hypotheses concerning the effects of hybridization on plant resistance. Individually marked plants were identified using morphological traits in the field and measurements of stipule and leaf pubescence were made and compared using Canonical Discriminant Function Analysis. DNA was extracted from the leaves of a sample of the marked plants and RAPD-PCR analysis was performed to establish the genetic status of parental and hybrid plants. RAPD band analysis generally verified the genetic status of parental plants. Hybrid plants were usually correctly identified in the field with a few exceptions. However, the hybrid plants were a heterogeneous group of plants made up of most plants that appear to be F₁s and a few plants that appear to be backcrosses to S. sericea. Morphological variables were useful for distinguishing S. sericea from S. eriocephala and hybrids, but were not as dependable in distinguishing between S. eriocephala and hybrids. We compared the densities of 11 herbivore species and the infection by a leaf rust pathogen (Melampsora sp.) on the leaves and stems of two parents and the hybrids in the field. We found support for the Additive hypothesis (3 species), the Dominance hypothesis (2 species) and the Hybrid Susceptibility hypothesis (7 species, 6 herbivores and the Melampsora rust). We found no evidence for the Hybrid Resistance hypothesis. Guild membership was not a good predictor of similar responses of species to hybrid versus parental plants. A Canonical Discriminant Function Analysis showed discrete separation of the taxa based on herbivore densities, illustrating different community structures on hybrid and parental plants. This study demonstrates the diversity of responses of phytophages in response to interspecific hybridization.     

Plant age, communication, and resistance to herbivores: young sagebrush plants are better emitters and receivers

Plants progress through a series of distinct stages during development, although the role of plant ontogeny in their defenses against herbivores is poorly understood. Recent work indicates that many plants activate systemic induced resistance after herbivore attack, although the relationship between resistance and ontogeny has not been a focus of this work. In addition, for sagebrush and a few other species, individuals near neighbors that experience simulated herbivory become more resistant to subsequent attack. Volatile, airborne cues are required for both systemic induced resistance among branches and for communication among individuals. We conducted experiments in stands of sagebrush of mixed ages to determine effects of plant age on volatile signaling between branches and individuals. Young and old control plants did not differ in levels of chewing damage that they experienced. Systemic induced resistance among branches was only observed for young plants. Young plants showed strong evidence of systemic resistance only if airflow was permitted among branches; plants with only vascular connections showed no systemic resistance. We also found evidence for volatile communication between individuals. For airborne communication, young plants were more effective emitters of cues as well as more responsive receivers of volatile cues.     

导入β—1,3—葡聚糖酶及几丁质酶基因的转基因可育油菜及其？…

利用农杆菌转化法,将组成型表达β－１,３－葡聚糖酶及必丁质酶基因的双价值植物表达载体ｐＢＬＧＣ转化优质甘蓝型油菜品种Ｈ１６５,并得到了抗卡那毒素（Ｋａｎａｍｙｃｉｎ,Ｋａｎ）的再生植株。我们对所得到的抗Ｋａｎ的再生苗进行了初步分子生物学检测,结果表明,在Ｋ１５（Ｋａｎ １５ｍｇ／Ｌ）培养基上的绿苗中有３０％为ＰＣＲ阳性植株,而在Ｋ２５（Ｋａｎ １０ｍｇ／Ｌ）培养基上的绿苗有５３％的阳性率。对部分Ｐ     

Leaf surface wetness in sorghum and resistance to shoot fly, Atherigona soccata: role of soil and plant water potentials

In experiments with potted plants, the relationships between soil matric potential, plant water potential and production of water droplets (leaf surface wetness) on the folded central whorl leaf of seedlings of sorghum genotypes that are either resistant or susceptible to shoot fly (Atherigona soccata) damage were investigated. Differences in soil matric potentials in the pots affected the plant water status, which in turn had profound effects on the production of water droplets on the central whorl leaf of the sorghum genotype susceptible to shoot fly. There was no consistent variation in the relationship between plant water potential and soil matric potential of resistant and susceptible sorghum genotypes. However, there was very little or practically no water droplets on the central whorl leaf of the resistant genotypes, indicating that the production of water droplets is not solely the result of internal water status of the plant. It is suggested that leaf surface wetness is genetically controlled and that an understanding of the mechanism by which water is transferred to the leaf surface will enhance breeding for resistance to shoot fly.     

Field screening maize germplasm for resistance and tolerance to western corn rootworms (Col.: Chrysomelidae)†

Abstract: In the hopes of lessening the current reliance on soil insecticides, developing a viable alternative for transgenic maize hybrids, and providing sustainable options for Europe, researchers recently have been developing novel maize lines that exhibit resistance and/or tolerance to corn rootworm larvae. Here we report the results of a 2‐year field experiment in a northern growing region assessing the resistance and tolerance of 10 experimental synthetic maize populations selected for varying levels of damage from western corn rootworm larvae, Diabrotica virgifera virgifera LeConte (Col.: Chrysomelidae) and four maize hybrids. Maize non‐preference, antibiosis and tolerance to rootworms was evaluated using previously established methods, including: the Iowa 1–6 root damage rating scale, root fresh weight, compensatory root growth ratings and adult rootworm emergence. Among the experimental synthetic maize populations, BS29‐11‐01 was the most susceptible, and had a mean root damage rating that was greater than the highly susceptible maize hybrid B37 × H84. This line also had the lowest mean root fresh weight and one of the lowest mean compensatory root growth ratings. In contrast, CRW8‐3 appeared to be tolerant to western corn rootworms, and had the lowest mean root damage rating, which was comparable with that of the non‐transgenic hybrid DeKalb^® 46‐26.     

Lack of correlation between constitutive and induced resistance to a herbivore in crucifer plants: real or flawed by experimental methods?

The correlation between constitutive and induced resistance to herbivores in plants has long been of interest to evolutionary biologists, and various approaches to determining levels of resistance have been used in this field of research. In this study, we examined the relationship between constitutive and induced resistance to the diamondback moth, Plutella xylostella (L.) (Lepidoptera: Plutellidae), in 11 closely related species of wild crucifers. We assessed the survival, development, and reproduction of the test insects and calculated their intrinsic rate of increase as an indicator of constitutive and induced resistance for the plants. We used larvae of P. xylostella and jasmonic acid as elicitors of the induced response. We failed to find a correlation between constitutive and induced resistance in these crucifer plants when the induction of resistance was initiated by either herbivory or jasmonic acid application. Analysis of the results suggests that the failure to detect a relationship between the two types of resistance could be caused by flaws in measuring constitutive resistance, which was apparently confounded with induced resistance. We discuss the difficulties and pitfalls in measuring constitutive resistance and ways to improve the methodology in investigating the relationships between constitutive and induced resistance in plants.     

Host-plant effects the expression of resistance to Bacillus thuringiensis kurstaki in Trichoplusia ni (Hubner): an important factor in resistance evolution

Pathogens are thought to exert strong selection on their hosts leading to increased host resistance. Bacillus thuringiensis kurstaki (Bkt) is a ubiquitous entomopathogen that has become the mainstay of nonchemical control of Lepidopteran pests and thus, the potential exists for the evolution of resistance in targeted host insects. We have studied the expression of Btk resistance in the cabbage looper, Trichoplusia ni (Hubner). For this generalist insect herbivore, three common host plants, tomato, pepper and cucumber, vary in suitability for larval growth and development. Here we show that the host plant also affects the overall toxicity of Btk, the relative expression of resistance between a resistant and a susceptible line and their F(1) reciprocal crosses, and importantly, the dominance of the resistance trait. This study demonstrates that tri-trophic interactions involving an insect, host plants and a pathogen have the potential to strongly influence the evolutionary response of an insect host to a pathogen.