Aphyocypris chinensis is a small cyprinid that is widely distributed in lowland areas of continental China, the Korean Peninsula, and the northwestern part of Kyushu, Japan. However, this species is severely threatened in Japan due to human impacts; thus, several facilities and citizen groups implemented captive breeding and reintroduction/reinforcement programs using several dozen founder fish collected from Tanushimaru, the location with the only known natural population, in the 1980s and 1990s. To determine the phylogenetic position and genetic authenticity of Japanese populations of A. chinensis, we conducted mtDNA and genome-wide SNP analyses using a total of 254 specimens from 31 wild and captive samples, including specimens of Chinese and Korean A. chinensis and the close relative A. kikuchii from Taiwan. The mtDNA divergence and phylogeny indicated that the haplotypes found exclusively in Japan were differentiated from the Chinese/Korean haplotypes (uncorrected p = 2.6% in the cytochrome b gene) to the same extent as they were from A. kikuchii haplotypes (p = 2.9%). Results from mtDNA sequences and 47-359 SNPs, obtained using the MIG-seq method with different parameter sets, revealed that the initial captive populations and an extinct wild population in Ukiha, adjacent to Tanushimaru, were genuine Japanese populations, whereas all extant captive and wild populations in Japan are hybrids between Japanese A. chinensis and A. kikuchii, or A. kikuchii itself. The details of the captive breeding and exchange programs, as well as evidence for the aquarium trade of A. kikuchii since the 1990s, strongly suggest that the mixture of A. kikuchii into Japanese populations occurred within the first several years of the establishment of captive populations in 1994. The present case of the highly probable extinction of genuine Japanese populations of A. chinensis emphasizes the importance of confirmation and management of the genetic authenticity of conservation-targeted species. Keywords Captive breeding • Endangered species • Ex situ conservation • Hybridization • MIG-seq • mtDNA cytochrome b JPN-1 (n=2) HFC16, KEE04 JPN-2 (4) LBM00,01, KEE04 JPN-3 (2) UK08 JPN-5 (1) UK07 JPN-4 (10) TNS08, CPR18, LBM09, KEE04 AB218688 TNS CK-1 (1) KOR07 CK-4 (2) KOR07 CK-6 (2) KOR07 CK-7 (1) KOR07 CK-8 (2) KOR07 CK-2 (2) CLN01 CK-3 (1) KOR07 CK-5 (1) KOR07 AF307452 CN KKC-1 (138) TNS08… JX184925 TW-Hualien KKC-2 (1) THL02 AP012123 Aquarium KKC-3 (1) THL02 CSC-2 (1) CSC02 CSC-1 (1) CSC02 AP011398 Yaoshanicus arcus AP011396 Nicholsicypris normalis JX311312 Pararasbora moltrechti 0.08

Spatial distributions of the biomass and species composition in submerged macrophytes in the southern basin of Lake Biwa were surveyed in September, 2002. Macrophyte samples were collected from 52 sites by SCUBA. The total biomass (dry weight) of and the vegetated area of macrophytes in the southern basin during the study period were estimated as 10735 ± 3030 t, and 43 km 2 , respectively. Potamogeton maackianus was dominant, and Hydrilla verticillata, Ceratophyllum demersum, Egeria densa and Myriophyllum spicatum were also abundant in the basin. Biomass of these five species accounted for 99% of the total macrophyte biomass. P. maackianus and C. demersum showed high niche overlap (Pianka's α = 0.71). Biomass of P. maackianus was positively correlated with the average transparency/water depth ratio (Spearman's ρ = 0.50), suggesting that P. maackianus tended to grow at the bottom under strong light. Biomass of C. demersum showed a positive correlation with transparency (ρ = 0.44). Biomass of E. densa was negatively correlated with the sediment diameter

We have developed a procedure to process echosounding data to map the distribution of submerged aquatic macrophytes in the southern basin of Lake Biwa, a water body that has a surface area of 52 km 2 and a mean depth of 4 m. Echosounding observations were made along 27 transect lines spaced at 500-m intervals on August 4 and September 2 and 30, 2003. Quantitative vegetation data including percent coverage, mean vegetation height, and percent vegetation infestation were directly determined using image data from the echosounder recorded digitally on videotape. Based on the image data from an echosounder, a regression model was developed for estimating biomass of submerged macrophytes. The regression model using the total echo strength as the explanatory variable could reliably estimate macrophyte biomass up to 300 g m −2 . Distribution maps of macrophyte height and biomass suggest that the recent summer decline of submerged macrophytes started earlier in shallow areas (<3 m of depth) than deep areas (>4 m) in the southern basin of Lake Biwa.

The molecular structures and pasting properties of four legume starches (white Dutch runner; WDR, adzuki bean, pea and common bean) were examined. The apparent amylose content (27 .0-30.0%) was 3.9-9.4% higher than the actual amylose content (17.6-25.5%) due to the high iodine affinity (IA) of the legume amylopectins. The crystalline type of pea starch was of Cb and those of the others were of the Cc. Adzuki bean starch had the highest maximum viscosity and breakdown. No breakdown was observed for pea starch. The legume amylopectins had IA of 1.32-2.28 g/100 g and number-average chain-length of 21-22. The phosphorus content of amylopectin varied be tween 13 and 820 ppm with WDR amylopectin having the highest content. All the legume amylo pectins had 1-4% of long-chain fraction, being consistent with their high IA. Compared with starches of cereal amylopectins, the legume amylopectins seemed to contain a relatively small amount of A-chain fraction. Distribution pattern for chains of degree of polymerization (DP) 6-8 was similar to those of potato and sweet potato amylopectins in which the amount of these chains decreased with an increase in DP. The legume amyloses had number-average DP of 820-1350 and comprised 2.4-4.7 chains per molecule, being similar to cereal amyloses. Among the legume amy loses, adzuki bean amylose was the largest and most branched molecule.

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