The Preliminary Study on DNA Barcoding of Mosses——A Case of Part of Genera of Meteoriaceae

We compared the performances of the candidate loci for moss DNA barcoding and the primers used in amplifying the loci. Primers for three coded loci (matK, rps4 and rbcL a) and four non coded loci (atpB rbcL, atpF H, psbK I and trnH psbA) of the chloroplast genome, one from the mitochondrial genome (nad5), and one from the nucleus genome (ITS2) were evaluated. Seventy four samples representing 14 species belonging to five genera of Trachypodoaceae (or Meteoriaceae) were screened. All primers for matK and a pair of primers for trnH psbA failed. Low successes were encountered with the primers for atpF H and psbK I. The primers for psbK I produced several bands and the PCR products of atpF H were difficult to sequence. The powers of the remaining six loci were compared using the variability, identification success and the resolutions. It was found that ITS2 is the most promising candidate for DNA barcoding for mosses. Among the chloroplast genes, atpB rbcL exhibited the highest resolution. Although trnH psbA is very variable, it is too short to be an ideal barcode alone. Combinations of chloroplast genes were also tried and Ps of both atpB rbcL+trnH psbA and rbcL a++trnH psbA were 64% using NJ method. More additions of loci did not increase the resolution. No barcoding gap exists for all these loci. Phylogenetic analyses were carried out prior to the DNA barcoding evaluation and some taxonomic problems do exist. This study exemplifies the necessity of correct species delimitation and the adoption of both plastid and nuclear loci in plant DNA barcoding.     

Physical and genetic mapping of safflower chloroplast DNA

Summary A physical map of safflower (Carthamus tinctorius L.) chloroplast DNA has been generated using SalI, PstI, KpnI and HindIII restriction endonucleases. The circular plastid genome (151 kbp) has the usual inverted repeat. Heterologous probes containing psbA, rbcL, atpA or rrnA structural genes mapped colinearly with spinach and other chloroplast genomes.     

Chloroplast gene for Mr 32000 polypeptide of photosystem II in Euglena gracilis is interrupted by four introns with conserved boundary sequences

The gene for the Mr 32000 herbicide binding polypeptide of photosystem II has previously been mapped to the 5 kbp EcoRI fragment Eco I of Euglena gracilis chloroplast DNA. The nucleotide sequence of 3324 bp of Eco I, containing the psbA locus, has been determined. This locus encodes a polypeptide of 345 amino acids which is co-linear with, and has 86% derived amino acid sequence homology to sequences derived from four higher plants chloroplast psbA loci. The Euglena psbA gene contains four introns of size 435, 443, 434, and 617 bp. The four introns have conserved boundary sequences of the type previously described in the Euglena chloroplast gene (rbcL) for the large subunit of ribulose-1,5-bisphosphate carboxylase (Koller et al., Cell 36, 545-553, 1984).     

Capsule formation and asymbiotic seed germination in some hybrids of Phalaenopsis,influenced by pollination season and capsule maturity

Physiology and Molecular Biology of Plants - We explored the influence of pollination season and maturity of capsule on post-pollination capsule formation and in vitro asymbiotic seed germination,...     

Chloroplast to chromoplast transition in tomato fruit: spectral confocal microscopy analyses of carotenoids and chlorophylls in isolated plastids and time-lapse recording on intact live tissue

Background and Aims

There are several studies suggesting that tomato (Solanum lycopersicum) chromoplasts arise from chloroplasts, but there is still no report showing the fluorescence of both chlorophylls and carotenoids in an intermediate plastid, and no video showing this transition phase.

Methods

Pigment fluorescence within individual plastids, isolated from tomato fruit using sucrose gradients, was observed at different ripening stages, and an in situ real-time recording of pigment fluorescence was performed on live tomato fruit slices.

Key results

At the mature green and red stages, homogenous fractions of chloroplasts and chromoplasts were obtained, respectively. At the breaker stage, spectral confocal microscopy showed that intermediate plastids contained both chlorophylls and carotenoids. Furthermore, an in situ real-time recording (a) showed that the chloroplast to chromoplast transition was synchronous for all plastids of a single cell; and (b) confirmed that all chromoplasts derived from pre-existing chloroplasts.

Conclusions

These results give details of the early steps of tomato chromoplast biogenesis from chloroplasts, with the formation of intermediate plastids containing both carotenoids and chlorophylls. They provide information at the sub-cellular level on the synchronism of plastid transition and pigment changes.     

Positioning of protein-coding genes on the soybean chloroplast genome

Seven major plastid protein encoding genes were positioned on the soybean chloroplast DNA by heterologous hybridization. These include the genes for the alpha, beta and epsilon subunits of the CF₁ component of ATP synthase (atpA, atpB and atpE respectively), for subunit III of the CF₀ component of ATP synthase (atpH), for the cytochrome f (cytF), for the ‘32 Kd’ thylakoid protein (psbA), and for the large subunit of ribulose-1,5-bisphosphate carboxylase-oxygenase (rbcL), all of which map in the large single copy region. The atpB, atpE and rbcL genes are located in the region adjacent to one of the segments of the inverted repeat. The genetic organization of the soybean chloroplast DNA is compared to that of other plastid genomes.     

中国秋海棠属 (秋海棠科) 植物的DNA条形码评价

秋海棠属植物种类繁多,形态变异多样,导致种类的系统放置混乱,近缘种类鉴定困难。利用DNA条形码实现物种快速准确的鉴定技术具有不受形态特征约束的优势,为秋海棠属植物的分类鉴定提供了新的方法。本研究选择4个DNA条形码候选片段（rbcL,matK,trnH psbA,ITS）对中国秋海棠属26种136个个体进行了分析。结果显示：叶绿体基因rbcL,matK和trnH psbA种内和种间变异小,对秋海棠属植物的鉴别能力有限;ITS/ITS2种内和种间变异大,在本研究中物种正确鉴定率达到100%/96%,可考虑作为秋海棠属DNA条形码鉴定的候选片段。研究结果支持中国植物条形码研究组建议将核基因ITS/ITS2纳入种子植物DNA条形码核心片段中的观点。