The Application of RAPD Markers in Diversity Detection and Variety Identification of Porphyra

RAPD (random amplified polymorphic DNA) markers were generated from filaments of 15 Porphyra lines representing four important groups, P. yezoensis, P. haitanensis, P. katadai var. hemiphylla and P. oligospermatangia . Among the total 69 fragments generated by 6 selected primers (among 50 primers), 67 appeared to be polymorphic (97.1%). Cluster analysis based on the RAPD results was performed. The 15 Porphyra lines were divided into 3 groups. This result was consistent with that from taxonomy analysis. A DNA fingerprinting based on 8 bands amplified with OPN_02 and OPJ_18 was constructed and might be used in Porphyra variety identification. Five specific RAPD fragments of 5 Porphyra lines were isolated and cloned into pGEM_T easy vector. These five RAPD fragments may be useful in germplasm identification and property protection of Porphyra .

AFLP Fingerprinting Analysis of Elite Hevea brasiliensis Germplasm

There are more than 6 000 clones of Hema brasiliensis Mull. Ary germplasm in the germplasm garden of Chinese National Key Biotechnology Laboratory for Tropical Crops and some of them are elite germplasm demonstrated by production and previous studies. AFLP (amplified fragment length polymorphism) fingerprinting analysis was performed on 25 clones (15 of Wickham clones and 10 of Amazon wild clones which possess phenotypes with high-yielding,/Iow-yielding, cold-tolerance/cold-sensitivity, oidium-resistance/oidium-sensitivity, tapping panel dryness (TPD) /healthy) respectively through a 377 DNA sequencer (P. E. Corp.) and PAGE electrophoresis results were analyzed by using GeneScan(TM) and Genotype(TM) Analysis software (P. E. Corp. The fragment profiles of different clones were obtained. Five hundred and eighteen fragments were generated by two primer combinations screened from 64 primer combinations and 511 fragments appeared to be polymorphic (98.6%). Genetic distance ranged from 0.25 to 0.81 between clones and ranged from 0.07 to 0.17 within RRIM600 clone. A specific 320 bp fragment of the oidium-resistant clones was found through genotype analysis. These results showed that AFLP fingerprints were highly reproducible and powerful and can be widely used in germplasm identification and genetic diversity analysis of Hema brasiliensis. In addition, based on the AFLP data, cluster analysis was performed. Cluster results showed that all the clones studied were almost clustered into a group one by one.

Assessing the Germplasm of Laminaria (Phaeophyceae) with Random Amplified PoLymorphic DNA (RAPD) Method

Eighteen gametophytes includingL. japonica, L. ochotensisandL. longissima, were verified with random amplified polymorphic DNA (RAPD) technique. Eighteen ten-base primers were chosen from 100 primers selected for final amplification test. Among the total of 205 bands amplified, 181 (88.3%) were polymorphic. The genetic distance among different strains ranged from 0.072 to 0.391. The dendrogram constructed by unweighted pair-group method with arithmetic (UPGMA) method showed that the female and male gametophytes of the same cell lines could be grouped in pairs respectively. It indicated that RAPD analysis could be used not only to distinguish different strains ofLaminaria, but also to distinguish male and female gametophyte within the same cell lines. There is ambiguous systematic relationship if judged merely by the present data. It seems that the use of RAPD marker is limited to elucidation of the phylogenetic relationship among the species ofLaminaria.

Identification of Porphyra lines using computerized DNA fingerprinting

RAPD (Randomly Amplified Polymorphic DNA) analysis was performed with filaments of 15 Porphyra lines representing four important groups (P. yezoensis, P. haitanensis, P. katadai var. Hemiphylla and P. digospermatangia). Eight stable and repeatable RAPD bands amplified with two primers, OPN-02 and OPJ-18, were selected for the construction of DNA fingerprinting. The RAPD results were scored based on the presence or absence of each of the 8 bands and then converted to computer language expressed with two digitals, 1 and 0, which represented the presence (numbered as 1) or absence (numbered as 0) of each band, respectively. Based on these results, a model DNA fingerprint and a computerized DNA fingerprint were constructed. In the constructed DNA fingerprint, each Porphyra line has its unique fingerprinting pattern and can be easily distinguished from each other. Later, a software, named as PhGI, was designed based on this DNA fingerprinting. It can be used in practical Porphyra line identification.

Research Progress of Polygonatum Germplasm Resources in China

Polygonatum is a traditional and precious Chinese medicine in China,as well as a traditional medicinal and food homologous material.It has a long history of cultivation and medicinal use.It not only has the effects of boosting qi and nourishing yin,nourishing the spleen and lungs,and tonifying the kidney,but also application value in food,health products,ornamental,cosmetics,etc.There are many varieties of Polygonatum resources all over China,which are easy to be mixed.There are three species of medicinal Polygonatum recorded in the 2015 edition of Pharmacopeia of the People s Republic of China.In order to sustainably use and fully develop Polygonatum medicinal resources,we conducted textual research on relevant data and research on the distribution of Polygonatum resources,germplasm identification,genetic diversity and breeding research,so as to provide a reference for the selection and breeding of polygonatum,artificial cultivation and the development and utilization of potential Polygonatum resources and for the solution of the contradiction between supply and demand in the Polygonatum market.

Development of EST-SSR primers and their practicability test for Laminaria

Laminaria （Saccharina） is one of the economic important seaweeds. Through sequence analysis of 4 099 ESTs from Laminaria （971 were generated from our own cDNA laboratories and 3 128 were downloaded from updated EST databases） with SSRIT software, two hundred and fifty-four SSRs in 206 ESTs were found. From the 254 SSRs, sixty-three SSR primer-pairs were designed. In order to test their practicability, the 63 primer-pairs were tested in commonly used SSR reaction conditions using 12 Laminara DNA samples as templates. The results show that 23 SSR primer-pairs gave good amplification patterns on most （more than 80%） of the 12 Laminara DNA templates. Genetic diversity study of 12 Laminaria lines, which were widely used in breeding and economic cultivation in China, was performed based on the obtained SSR data.

Structure Analysis of the Complete Mitochondrial Genome in Cultivation Variety ‘Rongfu’

In this report,complete mitochondrial genome sequences of Laminaria cultivation variety 'Rongfu' were obtained.The results showed the length of circular molecule of mtDNA was 37 638 bp(64.7% A+T),encoding three rRNAs(23S,16S and 5S),25 tRNAs,35 known mitochondrial proteins and 3 ORFs.Sequence alignment indicated its mtDNA genome was very similar to that of Laminaria japonica.Phylogenetic trees inferred from concatenated 30 mitochondrial genes showed that 'Rongfu',Laminaria japon-ica,Laminaria longipedalis,Laminaria diabolica,Laminaria religiosa and Laminaria ochotensis clustered together.In addition,compared with mitochondrial genome of L.japonica,'Rongfu' mtDNA lacked a non-coding region of 19 nucleotides,which was located between rRNA small subunit gene 3(rps3)and rRNA small subunit gene 9(rps9).Seven cultivation varieties of China were divided into two groups based on this non-coding region which was absent in 'Rongfu','Fujian' and 'Sanhai' while present in 'Ailunwan','Dongfang No.2','Dongfang No.3' and 'Zaohoucheng'.So this variation can be used in germplasm identification of cultivation variety.

Construction of DNA Fingerprint Using SSR Marker for Hybrid Rice Cultivars Approved by Hunan Province

[Objective] This study aimed to construct DNA fingerprint for hybrid rice cultivars those have been approved by Hunan Province. [Method] The primers which produced polymorphic and bright DNA bands were selected to construct the DNA molecular fingerprint map for 77 major hybrid rice cultivars approved by Hunan Province. [ Result] A total of 48 SSR primers were selected. Every cultivar had its unique fingerprint map so that the obtained data could identify differ- ent hybrid rice cultivars. [ Conclusion] This study made great contributions to the perfection of hybrid rice germplasm identification.

The use of RAPD analysis to identify 4 main medicinal plants in Vitex

Objective： To identify the plants in Vitex including Vitex negundo L. , V. negundo var. cannabifolia （Sieb. et Zuee. ） Hand. -Mazz. , V. trifolia L. and V. trifolia L. var. simplicifolia Cham.. Methods： Both intra- and inter-species relationships among these plants were analyzed by RAPD marker. Twenty-one samples collected from different locations in China were tested using 25 RAPD arbitrary 10- mer primers which were screened from 32 primers. Cluster analysis was conducted by Ward＇s minimumvariance method of SAS software. Results： A total of 224 bands were produced and 128 bands showed polymorphism among 21 samples. The level of polymorphism within species was 51.7%. The dendrogram constructed based on RAPD analysis showed that 21 samples can be placed in 2 groups at the level of SPRS value of 0. 3636. The first group included V. trifolia var. simplicifolia and V. trifolia. The other consisted of V. negundo and V. negundo var. cannabifolia.. At the level of SPRS value of 0. 1, 21 samples can be obviously divided into 3 groups. V. trifolia and V. trifolia var. simplicifolia were clustered into one group, V. negundo and V. negundo var. cannabifolia were separately clustered to 2 groups. Conclusion： RAPD analysis is in good agreement with the traditional plant taxonomic classification, and fundamentally identical with their origins and morphologie characteristics, which indicates that the genetic relationship among samples is related to their center of origin. These medicinal species in Vitex have their specific bands which are available to identify. These specific bands can be used as species-specific molecular markers of Fruetus Vitieis for the application of germplasm identification and classification.