[Directing construction of CRISPR/Cas9 vector of SmPAL1 in Salvia miltiorrhiza by target efficiency detection in vitro].

To construct CRISPR/Cas9 vectors for the editing of SmPAL1 in the phenylpropane metabolic pathway of Salvia miltiorrhiza, CIRSPR/Cas9 target sites of SmPAL1 were designed by online software. Its target efficiencies were detected in vitro by enzyme digestion and sequences with highly efficiency were constructed into CRISPR/Cas9 vectors. Three possible CRISPR target sequences (SmPAL1-g1, SmPAL1-g2, SmPAL1-g3) were designed and the enzyme digestion efficiencies were 53.3%, 76.6% and 10.0%. SmPAL1-g1 and SmPAL1-g2 were constructed into vector VK005-03 named as VK005-03-g1 and VK005-03-g2. The results of sequencing showed that the two CRISPR/Cas target sequences were all constructed into VK005-03. Here we first laid the foundation for the study of SmPAL1 and provided an effective strategy for the screening of sgRNA.

[Advances in study of plant type â ¢ polyketide synthases].

Polyketides are a large class of natural products with notable structural diversity and different biological activities. They have essential pharmacological value for human health. In plants, the enzymes responsible for the formation of phenolic metabolites backbone structures are collectively known as type Ⅲ polyketide synthases (PKSs), which are the key enzymes for the polyketides biosynthesis. The PKSs catalyze a series of condensation reactions of two-carbon acetate units with an acyl starter. A brief overview of this group of enzymes, including their reaction mechanisms, function modification, expression regulation, molecular evolution, and recent interesting findings are presented here.

[Construction of A Genetic Linkage Map in Salvia miltiorrhiza].

Objective: On the basis of previous studies, to construct a genetic map of Salvia miltiorrhiza by using EST-SSR primers, to build a platform for positioning important traits relating to genes, the cloning and molecular marker-assisted in breeding of new varieties of Salvia miltiorrhiza. Methods: A total of 411 EST-SSR primers were used for PCR amplification to screen polymorphic markers in F1 mapping population derived from a cross between Salvia miltiorrhiza, two parents of which were the cultivars of ZH74 and BH18. Combined with the molecular marker data of previous studies, Joinmap 4. 0 software was used for map integration. Results: 411EST-SSR primers were screened from two parents, a total of 328 pairs of primers were amplified stable products,164 pairs of polymorphic primers were obtained. Conclusion: A linkage map of Salvia miltiorrhiza with 150 marker high density genetic is constructed.

[Construction of the first genetic linkage map of Salvia miltiorrhiza Bge. using SSR, SRAP and ISSR markers].

The first genetic linkage map of Salvia miltiorrhiza was constructed in 94 F1 individuals from an intraspecific cross by using simple sequence repeat (SSR), sequence-related amplified polymorphism (SRAP) and inter-simple sequence repeat (ISSR) markers. A total of 93 marker loci in the linkage map, consisting of 53 SSR, 38 SRAP and 2 ISSR locus were made up of eight linkage groups, covered a total length of 400.1 cm with an average distance of 4.3 cm per marker. The length of linkage groups varied from 3.3 -132 cm and each of them included 2-23 markers, separately. The result will provide important basis for QTL mapping, map-based cloning and association studies for commercially important traits in S. miltiorrhiza.

[Development and molecular cytogenetic identification of 1RS.1BL Translocation lines derived from Triticale x Tritileymus].

In the progenies of octoploid Triticale Jinsong49 crossed with octoploid Tritileymus, a stable germplasm line named Shannong 030-1 was obtained. Its chromosome number was 42, and 21 bivalents could be observed at PMC MI. FISH analysis by using genomic DNAs from S. cereale and genomic DNAs from L. mollis as probes, respectively, showed that Shannong 030-1 contained a pair of translocation chromosome between rye and wheat, and had no genetic materials from L. mollis. C-banding analysis showed that it was translocation lines of 1RS.1BL. Resistance identification showed Shannong 030-1 was immune to powdery mildew.