Nedim Mutlu

Dicamba resistance: enlarging and preserving biotechnology-based weed management strategies.

The advent of biotechnology-derived, herbicide-resistant crops has revolutionized farming practices in many countries. Facile, highly effective, environmentally sound, and profitable weed control methods have been rapidly adopted by crop producers who value the benefits associated with biotechnology-derived weed management traits. But a rapid rise in the populations of several troublesome weeds that are tolerant or resistant to herbicides currently used in conjunction with herbicide-resistant crops may signify that the useful lifetime of these economically important weed management traits will be cut short. We describe the development of soybean and other broadleaf plant species resistant to dicamba, a widely used, inexpensive, and environmentally safe herbicide. The dicamba resistance technology will augment current herbicide resistance technologies and extend their effective lifetime. Attributes of both nuclear- and chloroplast-encoded dicamba resistance genes that affect the potency and expected durability of the herbicide resistance trait are examined.

A major QTL for common bacterial blight resistance derives from the common bean great northern landrace cultivar Montana No. 5

Knowledge of the evolutionary origin and sources of pest resistance genes will facilitate gene deployment and development of crop cultivars with durable resistance. Our objective was to determine the source of common bacterial blight (CBB) resistance in the common bean Great Northern Nebraska #1 (GN#1) and GN#1 Selection 27 (GN#1 Sel 27). Several great northern cultivars including GN#1, GN#1 Sel 27, and Montana No.5 (the female parent of the common x tepary bean interspecific population from which GN #1 and GN # 1 Sel 27 were derived) and known susceptible checks were evaluated for CBB reaction in field and greenhouse environments. These genotypes and CBB resistant and susceptible tepary bean including Tepary #4, the male parent and presumed contributor of CBB resistance toGN#1 and GN#1 Sel 27, were assayed for presence or absence of three SCAR markers tightly linked with independent QTLs conditioning CBB resistance. The parents and F2 of Montana No. 5/GN #1 Sel 27 and Montana No.5/Othello(CBB susceptible) were screened for CBB reaction and SCAR markers. CBB resistance in Montana No.5 was comparable to that of GN#1 and GN#1 Sel27. The SAP6 SCAR marker present in GN#1 and GN#1 Sel 27 was also present in Montana No.5, and it co-segregated (R2 =35%) with the CBB resistance in the Montana No.5/Othello F2 population. Although a few CBB resistant and susceptible transgressive segregants were found in the F2 of MontanaNo.5/GN #1 Sel 27 and later confirmed by F3 progeny tests, SAP6 SCAR marker was present in all progenies. None of the tepary bean specific CBB resistance-linked SCAR markers were present in GN#1, GN#1 Sel 27, or Montana No.5. A cluster analysis of 169 polymorphic PCR-based markers across three common bean and Tepary #4 indicated that GN#1, GN#1 Sel 27, and Montana No.5 were closely related, and not related at all with Tepary #4.Thus, these results clearly indicate Montana No.5, not Tepary #4, as the source of CBB resistance in GN#1 and GN#1 Sel 27.

Differential Pathogenicity of Xanthomonas campestris pv. phaseoli and X. fuscans subsp. fuscans Strains on Bean Genotypes with Common Blight Resistance

Both the common bacterial blight (CBB) pathogen (Xanthomonas campestris pv. phaseoli) and X. fuscans subsp. fuscans, agent of fuscous blight, cause indistinguishable symptoms in common bean, Phaseolus vulgaris. Yield losses can exceed 40%. Lack of information about the specificity between X. campestris pv. phaseoli strains and major quantitative trait loci (QTL) or alleles conferring resistance makes the task of identifying genetic changes in host-pathogen interactions and the grouping of bacterial strains difficult. This, in turn, affects the choice of pathogen isolates used for germplasm screening and complicates breeding for CBB resistance. Common bean host genotypes carrying various sources and levels of resistance to CBB were screened with 69 X. campestris pv. phaseoli and 15 X. fuscans subsp. fuscans strains from around the world. Differential pathogenicity of the CBB pathogen was identified on the 12 selected bean genotypes. The X. fuscans subsp. fuscans strains showed greater pathogenicity than X. campestris pv. phaseoli strains having the same origin. African strains were most pathogenic. The largest variation in pathogenicity came from X. campestris pv. phaseoli strains that originated in Caribbean and South American countries. Pathogenic variation was greater within X. campestris pv. phaseoli than within X. fuscans subsp. fuscans strains. Implications for breeding for CBB resistance are discussed.

AFLP and SRAP markers linked to the mj gene for root-knot nematode resistance in cucumber

Nematoides causadores de galhas (Meloidogyne spp.) em raizes de pepino Cucumis sativus L.) sao de ocorrencia mundial. Marcadores moleculares ligados ao gene mj que confere resistencia a M. javanica em pepino podem auxiliar na selecao de plantas em programas de melhoramento genetico. Cem AFLP (EcoRI-MseI) e 112 SRAP foram usados para a selecao de parentais resistentes e susceptiveis, por meio de polimorfismos, para o desenvolvimento de marcadores moleculares ligados ao gene mj. Entre 100 oligonucleotideos iniciadores para AFLP, 92 geraram fragmentos amplificados de DNA e dois produziram candidatos a marcadores (E-ATT/M-CAA e E-AAC/M-CTG). Os dois fragmentos amplificados foram clonados e sequenciados. Oligonucleotideos iniciadores sintetizados a partir das sequencias de nucleotideos obtidas nao produziram fragmentos polimorficos entre os parentais. Alem disso, as sequencias de nucleotideos nao contem sitios de restricoes ou delecoes que possam ser convertidos em marcadores CAPS ou SCAR. As duas sequencias de nucleotideos obtidas para os marcadores polimorficos AFLP foram usadas primeiramente para sintetizar os oligonucleotideos iniciadores especificos D1F, D1R, D17F e D17R. Oligonucleotideos iniciadores senso e antisenso para SRAP foram usados em combinacoes com os quatro oligonucleotideos iniciadores especificos para identificacao de polimorfismos entre os parentais. Dentre 112 combinacoes testadas, 11 geraram polimorfismos entre individuos parentais. O programa MapMaker Exp 3.0 foi usado para analise desses 11 marcadores. Foram identificados dois marcadores inseridos no gene mj nas distancias de 16.3 e 19.3 cM. Os resultados indicam que esses marcadores podem ser usados para o desenvolvimento de marcadores moleculares para o gene mj.

Elucidating genetic relationships, diversity and population structure among the Turkish female figs.

A collection of 96 female Turkish fig (Ficus carica L.) accessions was studied to elucidate genetic structure and estimate diversity and genetic similarity distribution among the female figs present in Turkish genetic resources, using 157 molecular genome markers including 129 sequence-related amplified polymorphisms, 21 random amplified polymorphic DNAs, and 7 simple-sequence repeats. The plant samples mainly included Turkish fig collections selected throughout the country over the course of a half-century. Neighbor-joining analysis revealed continuous dissimilarity range, and it was difficult to classify figs into distinct groups. The principle component analysis produced similar results. The analysis of molecular variance indicated that 95 and 93% of genetic variation were explained by within geographic origins and similar fruit rind color, respectively. Sub-structuring Bayesian analysis assigned the 96 female figs into four sub-populations, and indicated that they were highly related. The corrected allelic pairwise distances among the six geographic origins were less than 5%. This study suggests that geography- and color-based groups were not genetically distinct among the Turkish figs.

First report of Columbia root-knot nematode (Meloidogyne chitwoodi) in potato in Turkey.

Columbia root-knot nematode, Meloidogyne chitwoodi Golden et al., was identified from potatoes, Solanum tuberosum L., collected from Nigde Province, Turkey in September 2006. Seed potatoes are the most likely source for this introduction. The nematode is currently found to be infecting potatoes grown in the Netherlands, Portugal, Belgium, Germany, the United States, Mexico, South Africa, and Argentina. M. chitwoodi acquired a quarantine status in Europe (1) because of its potential to become established worldwide and its high damage probability. Some countries prohibit import of both seed and table stock potatoes originating in states known to harbor M. chitwoodi. Lesions on the potatoes had discrete brown coloration with white central spots in the outer 1 cm of the tuber flesh. Female nematode densities averaged 3 to 5 per cm2 of a potato section beneath the lesions. Nematodes were morphologically identified as M. chitwoodi based on the perineal pattern of mature females and the tail shape of juveniles. Using PCR-restriction fragment length polymorphism of the 18S region (3) and the mtDNA COII-16S rRNA region (2) and intergenic spacer region between the 5S and 18S genes (4), individual juveniles were identified as M. chitwoodi based on their restriction fragment patterns. To our knowledge, this is the first report of Columbia root-knot nematode infecting potatoes in Turkey. The distribution of this nematode in potato fields throughout Turkey should be determined. References: (1) L. J. M. F. Den Nijs et al. Nematology 6:303, 2004. (2) T. O. Powers and T. S. Harris. J. Nematol. 25:1, 1993. (3) T. O. Powers et al. J. Nematol. 37:226, 2005. (4) J. Wishart et al. Phytopathology 92:884, 2002.

Enhancing soybean photosynthetic CO2 assimilation using a cyanobacterial membrane protein, ictB ☆

Soybean C3 photosynthesis can suffer a severe loss in efficiency due to photorespiration and the lack of a carbon concentrating mechanism (CCM) such as those present in other plant species or cyanobacteria. Transgenic soybean (Glycine max cv. Thorne) plants constitutively expressing cyanobacterial ictB (inorganic carbon transporter B) gene were generated using Agrobacterium-mediated transformation. Although more recent data suggest that ictB does not actively transport HCO3-/CO2, there is nevertheless mounting evidence that transformation with this gene can increase higher plant photosynthesis. The hypothesis that expression of the ictB gene would improve photosynthesis, biomass production and seed yield in soybean was tested, in two independent replicated greenhouse and field trials. Results showed significant increases in photosynthetic CO2 uptake (Anet) and dry mass in transgenic relative to wild type (WT) control plants in both the greenhouse and field trials. Transgenic plants also showed increased photosynthetic rates and biomass production during a drought mimic study. The findings presented herein demonstrate that ictB, as a single-gene, contributes to enhancement in various yield parameters in a major commodity crop and point to the significant role that biotechnological approaches to increasing photosynthetic efficiency can play in helping to meet increased global demands for food.

Cry1Ac-mediated resistance to tomato leaf miner (Tuta absoluta) in tomato

Tomato leaf miner (Tuta absoluta) is a major pest of tomato in regions with hot climates such as South America and the Mediterranean. This insect feeds on almost every part of the plant and forms galleries while feeding on the plant’s inner tissues. Thus, it can cause plant death and is costly and difficult to control with chemical sprays. In this study, we transferred a modified Bacillus thuringiensis cry1Ac gene to tomato plants via Agrobacterium tumefaciens mediated transformation. Introduction of the cry1Ac gene to the tomato genome was confirmed with PCR and Southern blot analysis in 12 independent events. Insertion sites of the transgene in the tomato genome were determined with TAIL-PCR (thermal asymmetric interlaced polymerase chain reaction) for four selected transgenic lines. Cry1Ac gene expression was verified at both the transcriptional and translational levels, with RT-qPCR and Western blot analyses, respectively. Expression of the Cry1Ac protein in tomato resulted in T. absoluta mortality rates of 38–100% depending on transgenic line. In addition, gallery formation was reduced in 57–100% of the transgenic plants. Moreover, it was found that a single copy of the gene in the hemizygous condition is sufficient to confer tolerance to leaf miner. This is the first reported development of tomato plants resistant to T. absoluta. These transgenic plants are promising for development of commercial tomato cultivars resistant to leaf miner, which will limit the use of environmentally harmful chemicals for control of this pest.