Sequence-related amplified polymorphism (SRAP), a new marker system based on a simple PCR reaction: its application to mapping and gene tagging in Brassica

We developed a simple marker technique called sequence-related amplified polymorphism (SRAP) aimed for the amplification of open reading frames (ORFs). It is based on two-primer amplification. The primers are 17 or 18 nucleotides long and consist of the following elements. Core sequences, which are 13 to 14 bases long, where the first 10 or 11 bases starting at the 5′ end, are sequences of no specific constitution (”filler” sequences), followed by the sequence CCGG in the forward primer and AATT in the reverse primer. The core is followed by three selective nucleotides at the 3′ end. The filler sequences of the forward and reverse primers must be different from each other and can be 10 or 11 bases long. For the first five cycles the annealing temperature is set at 35°C. The following 35 cycles are run at 50°C. The amplified DNA fragments are separated by denaturing acrylamide gels and detected by autoradiography. We tested the marker technique in a series of recombinant inbred and doubled-haploid lines of Brassica oleracea L. After sequencing, approximately 45% of the gel-isolated bands matched known genes in the Genbank database. Twenty percent of the SRAP markers were co-dominant, which was demonstrated by sequencing. Construction of a linkage map revealed an even distribution of the SRAP markers in nine major linkage groups, not differing in this regard to AFLP markers. We successfully tagged the glucosinolate desaturation gene BoGLS-ALK with these markers. SRAPs were also easily amplified in other crops such as potato, rice, lettuce, Chinese cabbage (Brassica rapa L.), rapeseed (Brassica napus L.), garlic, apple, citrus, and celery. We also amplified cDNA isolated from different tissues of Chinese cabbage, allowing the fingerprinting of these sequences. Received: 3 November 2000 / Accepted 24 November 2000     

Wide-band tracheids in leaves of genera in Aizoaceae: the systematic occurrence of a novel cell type and its implications for the monophyly of the subfamily Ruschioideae

 Stem and leaf anatomical characters of succulent plants in families of the order Caryophyllales were examined to determine phylogenetic and evolutionary trends within these families. Wide-band tracheids, novel tracheids having wide secondary walls hypothesized to prevent cell collapse under water stress, were found in three families–Aizoaceae, Cactaceae, and Portulacaceae. Wide-band tracheids are hypothesized to be a unique adaptation to severe drought conditions prevalent in the areas of origin for these families. Species of 89 genera of Aizoaceae were examined for the occurrence of these tracheids. If present, wide-band tracheids occurred only in leaves. Of the five subfamilies examined, these tracheids were found only in the subfamily Ruschioideae, but not in the putative basal genera of the ruschioids. The presence of wide-band tracheids is an apomorphic character for genera of Ruschioideae with the exception of the basal genera, and also suggests that further characters are needed to understand the alliance of the Apatesia and Cleretum groups with other ruschioid groups. Received April 19, 2000 Accepted December 26, 2000     

藏北草地退化与生态服务功能价值损失评估——以申扎县为例

藏北草地是我国重要的畜牧业生产基地和生态安全屏障。基于遥感和GIS技术,利用1990、2000、2010年3期不同时相的TM、ETM+和CEBERS遥感影像,对申扎县草地资源退化状况进行了遥感监测,并采用生态经济学评估模型对草地生态系统8个方面的服务功能价值损失进行了评估测算。结果表明:1990—2010年,申扎县草地退化面积增加了47.40×10~4hm~2,生态系统服务功能价值损失高达5.20×10~8元;其中1990—2000年,草地退化较严重,该时段也是生态系统服务功能价值损失较多的时期;2000—2010年,草地退化趋势变缓。藏北草地提供生物量价值仅约占生态系统服务功能总价值的7.0%,草地生态服务功能远大于其提供的生物量价值,因此必须从生态服务功能的的理念出发去经营草地,从而实现草地的可持续发展。