DNA Marker Transmission and Linkage Analysis in Populations Derived from a Sugarcane (Saccharum spp.) x Erianthus arundinaceus Hybrid.

Introgression of Erianthus arundinaceus has been the focus of several sugarcane breeding programs in the world, because the species has desirable traits such as high biomass production, vigour, ratooning ability and good resistance to environmental stresses and disease. In this study four genetic maps were constructed for two intergeneric populations. The first population (BC1) was generated from a cross between an Erianthus/Saccharum hybrid YC96-40 and a commercial sugarcane variety CP84-1198. The second population (BC2) was generated from a cross between YCE01-116, a progeny of the BC1 cross and NJ57-416, a commercial sugarcane cultivar. Markers across both populations were generated using 35 AFLP and 23 SSR primer pairs. A total of 756 and 728 polymorphic markers were scored in the BC1 and BC2 populations, respectively. In the BC1 population, a higher proportion of markers was derived from the Erianthus ancestor than those from the Saccharum ancestor Badila. In the BC2 population, both the number and proportion of markers derived from Erianthus were approximately half of those in the BC1 population. Linkage analysis led to the construction of 38, 57, 36 and 47 linkage groups (LGs) for YC96-40, CP84-1198, YCE01-116, and NJ57-416, encompassing 116, 174, 97 and 159 markers (including single dose, double dose and bi-parental markers), respectively. These LGs could be further placed into four, five, five and six homology groups (HGs), respectively, based on information from multi-allelic SSR markers and repulsion phase linkages detected between LGs. Analysis of repulsion phase linkage indicated that Erianthus behaved like a true autopolyploid.

Transgenic sugarcane plants expressing high levels of modified cry1Ac provide effective control against stem borers in field trials.

To improve transgene expression level, we synthesized a truncated insecticidal gene m-cry1Ac by increasing its GC content from 37.4 to 54.8%, based on the codon usage pattern of sugarcane genes, and transferred it into two sugarcane cultivars (ROC16 and YT79-177) by microprojectile bombardment. The integration sites and expression pattern of the transgene were determined, respectively, by Southern, northern and western blot analyses. The transgenic sugarcane lines produced up to 50 ng Cry1Ac protein per mg soluble proteins, which was about fivefold higher than that produced by the partially modified s-cry1Ac (GC% = 47.5%). In greenhouse plant assay, about 62% of the transgenic lines exhibited excellent resistance to heavy infestation by stem borers. In field trials, the m-cry1Ac transgenic sugarcane lines expressing high levels of Cry1Ac were immune from insect attack. In contrast, expression of s-cry1Ac in transgenic sugarcane plants resulted in moderately decreased damages in internodes (0.4-1.7%) and stalks (13.3-26.7%) in comparison with the untransformed sugarcane controls, which showed about 4 and 26-40% damaged internodes and stalks, respectively. Significantly, these transgenic sugarcane lines with high levels of insect resistance showed similar agronomic and industrial traits as untransformed control plants. Taken together, the findings from this study indicate a promising potential of engineering an insect-resistant gene to tailor its protein expression levels in transgenic sugarcane to combat insect infestations.

GISH characterization of Erianthus arundinaceus chromosomes in three generations of sugarcane intergeneric hybrids.

Within Erianthus, a genus close to Saccharum, the species E. arundinaceus has the potential to contribute valuable traits to sugarcane, including adaptation to biotic and abiotic stresses and ratooning ability. Sugarcane breeders have tried for a long time to use Erianthus species in their breeding programs but until recently were constrained by a lack of fertile Saccharum x Erianthus hybrids. We report here for the first time the chromosome composition of fertile Saccharum officinarum x E. arundinaceus F1, BC1 (F1 x sugarcane cultivar), and BC2 (BC1 x sugarcane cultivar) hybrids. The F1 and BC2 resulted from n + n chromosome transmission, while the BC1 resulted from 2n + n transmission. In the BC1 clones, the number of E. arundinaceus chromosomes ranged from 21 to 30, and in the BC2 clones, the number ranged from 14 to 15, revealing cases of chromosome loss. No recombination events between Saccharum and Erianthus chromosomes were observed in either the BC1 or BC2 clones. The implications of these results for introgression of genes from E. arundinaceus in sugarcane breeding programs are discussed. We propose a strategy to identify the agronomic value of chromosomes from E. arundinaceus and to conduct targeted breeding based on this information.

[Effect of mutation of chemotaxis signal transduction gene cheA in Pseudomonas putida DLL-1 on its chemotaxis and methyl parathion biodegradation].

Pseudomonas putida DLL-1 is a high effective methyl parathion (MP) degrading strain, which is also of chemotaxis to MP. cheA, responsible for coding histidine kinase, plays an important role in bacterial chemotaxis signal transduction. In order to study the effect of bacterial chemotaxis on in-situ biodegradation of pesticides, cheA in chromosome of P. putida DLL-1 was mutated by gene-targeting and successfully obtained a MP-chemotaxis mutant DAK. The mutant DAK shows the same growth ability as wild-type DLL-1 in LB medium. The degrading rate of 50 mg/kg MP in non-sterile and sterile soil of chemotaxis mutant DAK is about 20%-30% lower than that of wild-type bacterial DLL-1. Lose of chemotaxis in DLL-1 would decline its degradation ability of MP. It was demonstrated that chemotaxis plays an important role in the in-situ biodegradation of pesticides.