Axel T. Lehrer

Genetic transformation with untranslatable coat protein gene of sugarcane yellow leaf virus reduces virus titers in sugarcane

Sugarcane yellow leaf syndrome, characterized by a yellowing of the leaf midrib followed by leaf necrosis and growth suppression, is caused by sugarcane yellow leaf virus (SCYLV). We produced SCYLV-resistant transgenic sugarcane from a susceptible cultivar (H62-4671) and determined the amount of virus present following inoculation. The transgenic plants were produced through biolistic bombardment of cell cultures with an untranslatable coat protein gene. Presence of the transgene in regenerated plants was confirmed using PCR and Southern blot analysis. The transgenic lines were inoculated by viruliferous aphids and the level of SCYLV in the plants was determined. Six out of nine transgenic lines had at least 103-fold lower virus titer than the non-transformed, susceptible parent line. This resistance level, as measured by virus titer and symptom development, was similar to that of a resistant cultivar (H78-4153). The selected SCYLV-resistant transgenic sugarcane lines will be available for integration of the resistance gene into other commercial cultivars and for quantification of viral effects on yield.

High incidence ofSugarcane yellow leaf virus (SCYLV) in sugar plantations and germplasm collections in Thailand

Sugarcane plantations and germplasm collections from across Thailand were tested in two surveys within the years 2000–2003 forSugarcane yellow leaf virus infection. Twenty-five to 100% of cultivars tested at each plantation/ germplasm collection were infected, among them those which had been imported from international breeding stations. Plantation management based on resistant cultivars or virus-free seed cane plantation practices is proposed.

Carbohydrate composition of sugarcane cultivars that are resistant or susceptible to Sugarcane yellow leaf virus

Sugarcane cultivars with a high (susceptible cultivars) and low (resistant cultivars) virus titer of Sugarcane yellow leaf virus were grown in the field. The carbohydrate composition in green leaf tops and in stems was determined. In RT-PCR of leaf extracts, susceptible cultivars had a high SCYLV-titer, whereas resistant cultivars had a very low titer. The cultivars differed in biomass yield, but these differences were not correlated with susceptibility. However, carbohydrate composition did have susceptibility-specific differences. Hexose levels were lower in green leaf tops and stalks of susceptible (strongly infected) cultivars than in those of resistant (weakly infected) cultivars. The stalks of susceptible cultivars also had less starch than those of resistant cultivars. Thus, the viral susceptibility (and infection) affected sugar metabolism. In addition, a positive correlation between hexose and starch in stems and between hexose and sucrose in green leaf tops was observed. The results from susceptible versus resistant cultivars were the opposite of those in the comparison between infected versus virus-free lines of the same cultivar. The breeding process apparently had unintentionally selected clones with modulated carbohydrate metabolism to avoid or compensate for the adverse effects of SCYLV infection.

Assessment of sucrose transporters, metabolites and sucrose phosphate synthase in different sugarcane tissues

The expression of sucrose-phosphate synthase II (SPSII) and sucrose transporters ShSUT1A and ShSUT4 were determined by RT-PCR and qRT-PCR in the sink and source leaves and in rind and pith of mature internodes of four high-yielding Hawaiian sugarcane cultivars. Expression of SPSII, ShSUT1A, and ShSUT4 was lower in pith than in rind, except in one cultivar, but else quite similar in the cultivars. The strong expression of transporter ShSUT4 in the rind of the internodes may hint to a special role of ShSUT4 in the rind. ShSUT4-expression in the sink and source leaves was similar in all four cultivars, whereas large differences were found for the expression of ShSUT1A and SPSII between the source and sink leaves and between the cultivars. The levels of sucrose precursors were doubled in source leaves compared to sink leaves, whereas they were higher in immature internode compared to mature internode. The role of sucrose transporters and SPSII in leaves and internodes is discussed, but the large differences, which were observed in the transcript levels of SPSII and sucrose transporters between some cultivars, although all the cultivars were similarly high-yielding cultivars, show that SPSII and SUT transcript levels cannot be used as indicators of high-yield cultivars.