Resistance to Bemisia tabaci in tomato wild relatives

Bemisia tabaci is one of the most threatening pests in agriculture, particularly in Solanaceous crops such as tomato and pepper that are cultivated in the open field. Pesticide application is often not effective and hazardous to humans and environment. The exploitation of plant natural defenses that are present in wild relatives of tomato, may offer a solution. To evaluate resistance parameters and to identify plant material with high levels of resistance, we screened a number of accessions of tomato wild relatives using three methods; a free-choice test in a screenhouse in Indonesia, a no-choice test with clip-on cages in a greenhouse and a leaf disc test in a climate-room in the Netherlands. Antibiosis resulting in low adult survival was the major component for resistance in tomato. However, other resistance component(s) may play a role as well. In some accessions there was a change in the resistance level over time. Several resistance parameters used in the different tests were well correlated. The best resistance source was an accession of Solanum galapagense, which had not been identified as being resistant in the past. This is of particular interest as this species is closely related to the cultivated tomato, which may facilitate introgression of the resistance component(s). Whitefly non-preference and resistance were associated with the presence of type IV trichomes. Other mechanisms might be involved since some accessions without type IV trichomes showed low nymphal density. The leaf disc test is a good in vitro alternative for the clip-on cage whitefly resistance screening, as shown by the high correlation between the results obtained with this test and the clip-on cage test. This offers breeders the possibility to carry out tests more efficientl

Regeneration of zygotic-like microspore-derived embryos suggests an important role for the suspensor in early embryo patterning

Plant science

Innovations in microspore embryogenesis in Indonesian hot pepper (Capsicum annuum L.) and Brassica napus L.

Hot pepper (Capsicum annuumL.) is the most important vegetable inIndonesia, but the yield is low, and the breeding programs are confined to the conventional methods and not efficient. To improve the efficiency of the breeding programs by speeding up the production of homozygous lines, studies were aimed at the introduction of haploid technology, which includes the regeneration and the production of doubled haploid plants from gametes. This technique is well developed in the model speciesBrassica napusL. via microspore culture. The results of various investigations involving both applied and fundamental aspects on microspore embryogenesis are presented in this thesis. The main results of the applied part deal with the development of an efficient shed-microspore culture protocol for the production of doubled haploid plants in Indonesian hot pepper (C. annuum), and its implementation under the local conditions of Indonesia. With regard to the more fundamental part, we presented for the first time an entirely new developmental pathway of embryogenesis including suspensor formation in microspore culture ofB. napuscv. Topas that mimics zygotic embryogenesis from early stages of development onwards. These results will have significant impact for practical application in hot pepper breeding programs as well as for further fundamental research on unraveling of early plant embryogenesis

Evaluation of crucial factors for implementing shed-microspore culture of Indonesian hot pepper (Capsicum annuum L.) cultivars

A shed-microspore culture protocol was developed in Wageningen for producing doubled haploid plants in several genotypes of Indonesian hot pepper (Capsicum annuum L.). For transfer of technology to Indonesia, three factors were studied that appeared crucial for successful implementation in practice. First, application in the culture medium of a combination of the antibiotics timentin and rifampicin at the concentrations of 200 and 10 mg/l, respectively, prevented bacterial contamination from the donor explants. Second, in vitro application of colchicine (100 ¿M) during the first week of culture was highly effective in increasing the percentage of doubled haploid plants. Third, a comparative analysis of the ploidy level of plants regenerated from shed-microspore-derived embryos using chloroplast counts in guard cells of leaf stomata and flow cytometric measurement of leaf nuclear DNA content, revealed that the first procedure is a reliable and an easy to use method for ploidy determination with hot peppe

Successful development of a shed-microspore culture protocol for doubled haploid production in Indonesian hot pepper (Capsicum annuum L.)

Various systems of anther and microspore cultures were studied to establish an efficient doubled haploid production method for Indonesian hot pepper (Capsicum annuum L.). A shed-microspore culture protocol was developed which outperformed all the previously reported methods of haploid production in pepper. The critical factors of the protocol are: selection of flower buds with more than 50% late unicellular microspores, a 1 day 4°C pretreatment of the buds, followed by culture of the anthers in double-layer medium system for 1 week at 9°C and thereafter at 28°C in continuous darkness. The medium contained Nitsch components and 2% maltose, with 1% activated charcoal in the solid under layer and 2.5 ¿M zeatin and 5 ¿M indole-3-acetic acid in the liquid upper layer. All the ten genotypes of hot pepper tested, responded to this protocol. The best genotypes produced four to seven plants per original flower bud. This protocol can be used as a potential tool for producing doubled haploid plants for hot pepper breeding

The Bemisia tabaci species complex: Additions from different parts of the world

Bemisia tabaci is one of the most threatening pests in many crops. We sequenced part of the mitochondrial cytochrome oxidase I gene from fifty whitefly populations collected in Indonesia, Thailand, India and China. Nineteen unique sequences (haplotypes) of the cytochrome oxidase I were identified in these populations. They were combined with sequences available in databases, resulting in a total of 407 haplotypes and analyzed together with nine outgroup accessions. A phylogenetic tree was calculated using the maximum likelihood method. The tree showed that all groups that were found in previous studies were also present in our study. Additionally, seven new groups were identified based on the new haplotypes. Most B. tabaci haplotypes grouped based on their geographical origin. Two groups were found to have a worldwide distribution. Our results indicate that our knowledge on the species complex around B. tabaci is still far from complete

The Bemisia tabaci species complex: Additions from different parts of the world

Bemisia tabaci is one of the most threatening pests in many crops. We sequenced part of the mitochondrial cytochrome oxidase I gene from fifty whitefly populations collected in Indonesia, Thailand, India and China. Nineteen unique sequences (haplotypes) of the cytochrome oxidase I were identified in these populations. They were combined with sequences available in databases, resulting in a total of 407 haplotypes and analyzed together with nine outgroup accessions. A phylogenetic tree was calculated using the maximum likelihood method. The tree showed that all groups that were found in previous studies were also present in our study. Additionally, seven new groups were identified based on the new haplotypes. Most B. tabaci haplotypes grouped based on their geographical origin. Two groups were found to have a worldwide distribution. Our results indicate that our knowledge on the species complex around B. tabaci is still far from complete

Resistance to Bemisia tabaci in tomato wild relatives

Bemisia tabaci is one of the most threatening pests in agriculture, particularly in Solanaceous crops such as tomato and pepper that are cultivated in the open field. Pesticide application is often not effective and hazardous to humans and environment. The exploitation of plant natural defenses that are present in wild relatives of tomato, may offer a solution. To evaluate resistance parameters and to identify plant material with high levels of resistance, we screened a number of accessions of tomato wild relatives using three methods; a free-choice test in a screenhouse in Indonesia, a no-choice test with clip-on cages in a greenhouse and a leaf disc test in a climate-room in the Netherlands. Antibiosis resulting in low adult survival was the major component for resistance in tomato. However, other resistance component(s) may play a role as well. In some accessions there was a change in the resistance level over time. Several resistance parameters used in the different tests were well correlated. The best resistance source was an accession of Solanum galapagense, which had not been identified as being resistant in the past. This is of particular interest as this species is closely related to the cultivated tomato, which may facilitate introgression of the resistance component(s). Whitefly non-preference and resistance were associated with the presence of type IV trichomes. Other mechanisms might be involved since some accessions without type IV trichomes showed low nymphal density. The leaf disc test is a good in vitro alternative for the clip-on cage whitefly resistance screening, as shown by the high correlation between the results obtained with this test and the clip-on cage test. This offers breeders the possibility to carry out tests more efficientl

Identification of silverleaf whitefly resistance in pepper

Whitefly is economically one of the most threatening pests of pepper worldwide, which is mainly caused by its ability to transmit many different viruses. In this research, we characterized pepper germplasm to identify whitefly-resistant accessions that will form the basis for future resistance breeding. Forty-four pepper accessions representing four species (Capsicum annuum, C. frutescens, C. chinense, C. baccatum) were screened for resistance to whiteflies. Screening parameters were adult survival (AS) and oviposition rate (OR) in a no-choice test and whitefly, egg and nymphal density in free-choice tests. To combine parameters in free-choice tests, a plant resistance value was calculated. The results show that AS and OR were significantly different among accessions and were positively correlated, which was also the case for the parameters in the free-choice tests. Accessions identified as highly resistant in no-choice and free-choice tests generally were C. annuum. Whitefly density and OR correlated positively with trichome density and negatively with cuticle thickness of leaves

Regeneration of zygotic-like microspore-derived embryos suggests an important role for the suspensor in early embryo patterning

The inaccessibility of the zygote and proembryos of angiosperms within the surrounding maternal and filial tissues has hampered studies on early plant embryogenesis. Somatic and gametophytic embryo cultures are often used as alternative systems for molecular and biochemical studies on early embryogenesis, but are not widely used in developmental studies due to differences in the early cell division patterns with seed embryos. A new Brassica napus microspore embryo culture system, wherein embryogenesis highly mimics zygotic embryo development, is reported here. In this new system, the donor microspore first divides transversely to form a filamentous structure, from which the distal cell forms the embryo proper, while the lower part resembles the suspensor. In conventional microspore embryogenesis, the microspore divides randomly to form an embryonic mass that after a while establishes a protoderm and subsequently shows delayed histodifferentiation. In contrast, the embryo proper of filament-bearing microspore-derived embryos undergoes the same ordered pattern of cell division and early histodifferentiation as in the zygotic embryo. This observation suggests an important role for the suspensor in early zygotic embryo patterning and histodifferentiation. This is the first in vitro system wherein single differentiated cells in culture can efficiently regenerate embryos that are morphologically comparable to zygotic embryos. The system provides a powerful in vitro tool for studying the diverse developmental processes that take place during the early stages of plant embryogenesis