SUMMARYCassava (Manihot esculenta Crantz) root yield measured as fresh weight (hereafter root yield) is declining in much of Asia and Africa. The current study aimed to identify quantitative trait loci (QTL) underlying both root and starch fresh weights in F1 cassava. Eight QTL were associated with root yield, underlying 12·9–40·0% of the phenotypic variation (PVE). Nine QTL were associated with starch content, underlying 11·3–27·3% of the PVE. Quantitative trait loci were identified from four different environments that encompassed two locations and 3 years. Consistent QTL for root yield, YLD5_R11 and YLD8_L09 on linkage group (LG) 16, were detected across years and locations. Quantitative trait loci for starch content, ST3_R09, ST6_R10 and ST7_R11 on LG 11, were found across 3 years. Co-localization of QTL for both traits with positive correlation was detected between YLD3_R10 and ST5_R10 on LG 9. Candidate genes within the QTL that were consistent across multiple environments were identified based on cassava genome sequences. Genes predicted to encode for glycosyl hydrolases, uridine 5’-diphospho-(UDP)-glucuronosyl transferases and UDP-glucosyl transferases were found among the 44 genes located within the region containing the QTL controlling starch content. Sixteen genes predicted to encode proteins that were possibly associated with root yield were identified. The QTL controlling root yield and starch content in the current study will be useful for molecular breeding of cassava through marker-assisted selection. The identification of candidate genes underlying both traits will be useful both as markers and for gene expression studies.

In this study, simple sequence repeats (SSRs) specific to marigold were developed using the inter-SSR technique and a SSR-enriched genomic DNA library. In addition, SSRs derived from sunflower (Helianthus annuus) were also tested for transferability to marigold. In total, 38 polymorphic markers with 112 observed alleles were identified in 20 African marigolds (Tagetes erecta L.) consisting of 14 commercial varieties and six Thai landraces, and six French marigolds (Tagetes patula L.). The number of alleles per locus ranged from 2 to 7. The averages of expected and observed heterozygosities were 0.48 and 0.32, respectively. Polymorphic information content values ranged from 0.10 to 0.71, and resolving power (R p ) values ranged from 0.23 to 2.77. The SSRs were successfully applied to the differentiation of the 26 marigold samples into clusters of African commercial varieties, Thai landraces and French marigold. The genetic relationship analysis revealed that the African commercial varieties were more closely related to the Thai landraces than to the French marigold. The results of the study indicate that the SSRs developed are effective for genetic diversity analysis, species classification and individual identification.

ABSTRACT. We developed molecular markers for cassava based on the microsatellite-amplified fragment length polymorphism (M-AFLP) technique. Twenty primer pairs were developed and used for the analysis of 48 samples of Manihot species, consisting of M. esculenta (33), M. esculenta ssp flabellifolia (3), M. chlorosticta (3), M. carthaginensis (3), M. filamentosa (3), and M. tristis (3). Nine microsatellite loci that were polymorphic among these Manihot species were identified, giving 32 polymorphic alleles and from two to seven alleles per locus. Unbiased and direct count heterozygosity varied from 0.0233 to 0.7924 and 0.0000 to 0.7083, respectively. There was significant deviation (P < 0.05) from Hardy-Weinberg equilibrium at five loci. Genotypic data from the Manihot species were subjected to genetic diversity analysis. We found that M. chlorosticta and M. esculenta ssp flabellifolia were the closest populations, while M. filamentosa and M. esculenta ssp flabellifolia were the most divergent. Considering within M. esculenta, the samples from Nigeria and Fiji were the most closely related, while those from Venezuela and of unknown origin were the most divergent. We conclude that the M-AFLP technique is an effective method for generating microsatellite markers that are useful for genetic diversity analysis in Manihot species.

Relative real-time reverse transcription quantitative polymerase chain reaction (RT-qPCR) is a wellestablished method for the precise quantification of gene expression. For accurate relative real-time RTqPCR analysis, validation of the expression of an appropriate reference gene is required. In this study, the expression of six commonly used reference genes, namely 40S ribosomal protein (40S), actin (ACT), cyclophilin C (CYCC), EF-1 alpha (EF1), TATA box binding protein (TBP) and polyubiquitin (UBI) was investigated in leaf and root samples of cassava obtained at 6, 9 and 12 months after planting (MAP). A transcript stability analysis was undertaken in two different varieties of cassava, namely Huay Bong 60 which has high cyanogenic potential (CN) and Hanatee which has low CN. The results reveal that TBP was the most stable reference gene for expression studies. This information was applied to an analysis of linamarase and α-hydroxynitrile lyase gene expression in samples from six low and six high CN cassava plants collected at 6, 9 and 12 MAP. The results indicate that at 6 MAP, the linamarase transcript from leaf of the high CN group was significantly increased, and the α-hydroxynitrile lyase transcript was significantly increased at 12 MAP.

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