ABSTRACT
PCR clamping by locked nucleic acid (LNA) oligonucleotides is an effective technique for selectively amplifying the community SSU rRNA genes of plant-associated bacteria. However, the original primer set often shows low amplification efficiency. In order to improve this efficiency, new primers were designed at positions to compete with LNA oligonucleotides. Three new sets displayed higher amplification efficiencies than the original; however, efficiency varied among the primer sets. Two new sets appeared to be available in consideration of bacterial profiles by next-generation sequencing. One new set, KU63f and KU1494r, may be applicable to the selective gene amplification of plant-associated bacteria.
Subject(s)
Bacteria/genetics , Bacterial Typing Techniques/methods , Microbiota/genetics , Oligonucleotides/genetics , Plants/microbiology , Polymerase Chain Reaction , Ribosome Subunits, Small, Bacterial/genetics , Bacteria/classification , Bacteria/isolation & purification , DNA Primers/genetics , DNA, Bacterial/genetics , High-Throughput Nucleotide Sequencing , Mitochondria/genetics , Plants/genetics , Plastids/genetics , Sequence Analysis, DNAABSTRACT
In order to study the effect of fertilization on the qualities of sweet pepper(Capsicum annul L) cultured in greenhouse, the experiment was carried out by drop irrigation. The results showed that the most sensitive organ of sweet pepper for nitrate was lateral branch, in which the nitrate content was highest. The nitrate content in leaf was lower than that in lateral branch, and in fruit, the nitrate content was the lowest. High fertilization of N, P and K had no negative influence on content of nitrate and total soluble sugars in fruit. The content of soluble sugars in fruit were about 50%-55% under all treatment and higher than that in leaf and lateral branch. Decreasing fertilization would cause increasing of the inosital and sucrose contents. Glucose and fructose were the main soluble sugars in fruit. The metabolism of carbohydrates was weakened by high fertilization, and it expressed as restraining the accumulation of starch content in leaf. The content of starch was the highest in fruit. The lipid content were exceed 3-4 times in the leaf than in the lateral branch and fruit, and perhaps, it was due to the higher content of chlorophyll in the leaf.
