Breeding of a practical rice line ‘TJTT8’ for phytoextraction of cadmium contamination in paddy fields

ABSTRACT

Previously, we showed that qCdp7, an allele identified in the high-Cd-accumulating indica rice variety ‘Jarjan,’ is associated with effective phytoextraction of Cd from paddy soils. However, ‘Jarjan’ may not be practical for phytoextraction because it is susceptible to seed shattering and culm lodging, which are unfavorable traits for mechanical rice harvesting. In this study, to develop a practical rice line for phytoextraction, we introduced the qCdp7 allele into ‘Tachisugata,’ a rice variety with a nonshattering habit and lodging resistance, to produce a new high-Cd-accumulating rice line designated ‘TJTT8.’ This line inherited high-Cd accumulation and brown pericarps from ‘Jarjan’ and a nonshattering habit and lodging resistance from ‘Tachisugata;’ all of these traits are necessary for rice intended for Cd phytoextraction in Japan. Backcross inbred lines (BILs) were produced by two backcrosses to ‘Tachisugata’ after a cross between ‘Jarjan’ and ‘Tachisugata.’ ‘TJTT8’ was selected from the BILs by means of marker-assisted selection and phenotypic evaluation. When ‘TJTT8,’ the parents, and ‘Cho-ko-koku’ which is a high-Cd-accumulating indica variety were cultivated in Cd-contaminated paddy fields in four locations in Japan, ‘TJTT8’ exhibited lodging resistance and shattering resistance that were higher than those of ‘Jarjan’ and ‘Cho-ko-koku’ and equivalent to those of ‘Tachisugata.’ ‘TJTT8’ accumulated Cd in the aerial parts of the plants at concentrations ranging from 6.5 to 22.7 mg m^?2: it showed significantly higher Cd accumulation than ‘Tachisugata’ and was equivalent to ‘Jarjan’ and slightly superior to ‘Cho-ko-koku.’ Soil Cd concentration was estimated to have been reduced by 8.7–33.6% based on the amount of Cd accumulation in the aerial parts of the plants. Thus, we succeeded in using the qCdp7 allele to produce a practical rice line for Cd phytoextraction by improving several agronomic traits for compatibility with Japanese cultivation systems.     

Cardiac remodeling and angiotensin II-forming enzyme activity of the left ventricle in hamsters with chronic pressure overload induced by ascending aortic stenosis

Cardiac remodeling and angiotensin II-forming enzyme activity of the left ventricle on chronic pressure overload were studied in male Syrian hamsters, whose chymase activity is similar to that of dogs. Pressure overload was achieved by banding at the ascending aorta (aortic stenosis). Echocardiography, histological analysis, and analysis of cardiac angiotensin-converting enzyme and chymase-like activities were performed. At 10 weeks after banding, concentric hypertrophy of the left ventricle was evident. At 20 weeks after banding, the ventricular weight-to-body ratio, cardiac fibrosis, and cardiac chymase-like activity were significantly increased, while cardiac angiotensin-converting enzyme activity was significantly decreased. This suggests that cardiac chymase, compared with cardiac angiotensin-converting enzyme, was activated against the chronic pressure overload and was responsible for the cardiac remodeling through the formation of angiotensin II. Considering the utility of the rodents, the interspecies similarity of the Ang II-forming pathway, and the effect of chymase in the hamsters, the present model is considered useful for studies evaluating the effect of Ang II and chymase in the canine heart with chronic pressure overload.     

Non‐invasive sampling technique for DNA extraction from captive Japanese Crested Ibis on Sado Island

The Japanese Crested Ibis Nipponia nippon is a critically threatened bird. The post‐hatch eggs of the current captive population of this species on Sado Island have been stored at room temperature for the long‐term. In this study, we investigated the suitability of the vascularized chorioallantois membrane from the eggs as a non‐invasive DNA source. Using microsatellite loci developed for the Japanese Crested Ibis, we performed three experiments for comparison of genotypes obtained among DNA. First, DNA from five different sites of the identical membrane showed the same genotypes at either of two loci examined. Second, DNA from the membrane of each full‐sibling birds and blood of their parents showed the genotypes that were consistent with Mendelian parent–offspring relationships at any of eight loci examined. Third, DNA from the membrane and blood of the same bird showed the matched genotypes at any of eight loci examined. These results indicate that the vascularized chorioallantois membrane from post‐hatch eggs stored at room temperature for the long‐ term can be used as a reliable DNA source of offspring that had hatched from the egg. This study will promote a molecular genetics study on genetic diversity of the current captive Japanese Crested Ibis population on Sado Island.     

An insertion/deletion variant of a thymine base in exon 2 of the porcine beta 3‐adrenergic receptor gene associated with loin eye muscle area

An insertion/deletion variant of a thymine base (T5 and T6) in exon 2 of porcine beta 3‐adrenergic receptor (ADRB3) gene has been described. In the current study, we made an association study between the ADRB3 polymorphisms and production traits in 735 Duroc pigs. The allele frequencies for the T5 and T6 alleles in our study population were 0.433 and 0.567, respectively. Any associations between ADRB3 genotype and average daily weight gain during test period, or backfat thickness and intramuscular fat content were not detected in either sex. However the size of the loin eye muscle area (EMA) was significantly associated with ADRB3 genotypes in gilts. T6‐homozygous gilts had a higher mean of EMA (40.6 ± 0.6 cm²) than T5‐homozygous (38.1 ± 0.4 cm², P = 0.002) and heterozygous (38.8 ± 0.3 cm², P = 0.034) gilts. This association was not detected in males. In addition, a multiple traits animal model best linear unbiased predictor (BLUP) analysis revealed that the T6‐homozygous genotype had positive effects on breeding value of EMA. Accordingly, we suggest that ADRB3 polymorphism has the potential to be an important genetic marker for prediction of EMA in Duroc pigs.     

Effects of nitrogen fertilizer and harvesting stage on photosynthetic pigments and phytol contents of Italian ryegrass silage

This study aimed to investigate the effects of nitrogen (N) fertilizer application and harvesting stage on the contents of chlorophyll, phytol and carotenoids (β‐carotene and lutein) in Italian ryegrass herbage before and after ensiling, and the extent of phytol preservation after ensiling. Three rates of N fertilizer (0, 60 and 120 kg N/ha) were applied by top‐dressing as an additional fertilizer. The herbage harvested at booting stage (27 weeks of age) or heading stage (29 weeks of age) were wilted for 1 day, then ensiled for 60 days using a small‐scale pouch system. In the pre‐ensiled herbages, increasing N fertilizer application increased the contents of crude protein and photosynthetic pigments, and these contents were also higher at the booting stage compared with the heading stage. In the silage, increasing N fertilizer application also increased the contents of crude protein, the photosynthetic pigments and their derivatives (pheophytin and pheophorbide), while harvesting stage did not affect the contents of β‐carotene, chlorophylls or pheophorbide. Nitrogen fertilizer application and early harvesting of herbage increased lutein and phytol contents in Italian ryegrass silage. Lutein and phytol in Italian ryegrass herbage are indicated to be well preserved during ensiling.     

Measurement of Carbonic Anhydrase I and II Isozymes in Feces as a Marker of Occult Blood in Horses with Intestinal Tract Bleeding

Although endoscopy is the definitive diagnostic method for the detection of colonic ulcers, the equipment required for performing the test is costly and difficult to use. Therefore, a simple cost-effective and reliable screening test for intestinal tract bleeding is needed. To this end, we measured carbonic anhydrase isozymes (CA-I and CA-II) originating from erythrocytes by ELISA in order to determine if they could be used as markers of occult blood in feces. For fecal extract preparation, 2 g of feces were mixed with 4 ml of 0.01 M Tris-HCl (pH 8.0) containing 0.01% thimerosal. The concentrations of CA-I and CA-II in the fecal samples of 13 clinically normal racehorses were found to be 30.0 ± 10.0 and 34.0 ± 13.0 ng/ml, respectively. Increased concentrations of CA-I were detected in the fecal samples of 5 horses after blood administration; however, no increase was observed in CA-II. The concentrations of CA-I and CA-II in the fecal samples of 88 racehorses with clinical signs of equine gastric ulcer syndrome (EGUS) were 115.3 ± 79.0 and 41.0 ± 42.0 ng/ml, respectively. Thus, our results indicate that CA isozymes can be useful as markers of occult blood in the fecal samples of horses with intestinal tract bleeding.     

Different effects on the inflammatory lesions in the lacrimal and salivary glands after neonatal thymectomy in IQI/Jic mice, a model for Sjögren's syndrome

Thymectomy on day 3 after birth (D3Tx) is understood to eliminate CD4+CD25+ regulatory T cells (Treg) from the peripheral T cell repertoire in rodents, leading to the activation of autoreactive T cells. Herein, D3Tx was performed in IQI/Jic mice, a model for Sj?gren's syndrome characterized by autoimmune infiltrations into the lacrimal and salivary glands. At the age of 16 weeks, very severe lesions were observed in lacrimal tissues from thymectomized mice, suggesting that Treg preserve their immunoregulatory function in young IQI/Jic mice. In contrast, salivary lesions were comparable in the D3Tx and control groups, raising the possibility that either salivary-specific Treg escaped elimination in thymectomized mice or spontaneous lesions in IQI/Jic mice developed independently of the tolerance through Treg.     

Histopathological, immunohistochemical and ultrastructural studies of a renal mesenchymal tumor in a young beagle dog

A 15-month-old male beagle dog used in a toxicity study had a primary renal mesenchymal tumor. Macroscopically, the tumor was a gray-white mass which was found in the right kidney, and extended from the capsule to a position slightly compressing the medulla. Microscopically, most of the tumor cells showed a myxoid pattern, in which the matrix was positive for alcian blue staining. In the other parts of the tumor, a fascicular and wavy pattern was observed, and the matrix was full of collagen fibrils. Immunohistochemically, tumor cells were positive for vimentin and fibronectin, and negative for cytokeratin, desmin, α-smooth muscle actin, Von Willebrand factor, cyclooxigenase-2 and myelin basic protein. As a result, we diagnosed this case to be a renal mesenchymal tumor. Based on the microscopic findings, interstitial characteristics and immunohistochemical features, the present case was classified as a congenital mesoblastic tumor.     

Association of an SNP marker in exon 24 of a class 3 phosphoinositide‐3‐kinase (PIK3C3) gene with production traits in Duroc pigs

A C?T single nucleotide polymorphism (SNP) on exon 24 of the porcine class 3 phosphoinositide‐3‐kinase (PIK3C3) gene is considered a possible genetic marker for selecting backfat (BF) thickness and carcass fat, although only one study has published results on its effects by performing experiments on a single resource family. We analyzed the association of this PIK3C3 polymorphism with production traits in 739 Duroc pigs. The C allele frequency was 67.9% in our study population. PIK3C3 polymorphism showed significant effects on average daily weight gain (ADG), BF thickness, intermuscular fat content (IMF), and the size of the loin eye muscle area (EMA). The C alleles increased ADG, BF and IMF, and decreased EMA. The predicted differences in traits between the homozygous pigs of the C and T alleles were 40 g/day for DG, 1.2 mm for BF, 0.44% for IMF, and 1.6 cm² for EMA. Furthermore, the statistical models for estimating the breeding values of each trait had lower Akaike's information criterion values when adding PIK3C3 genotype information. We therefore confirmed that the polymorphism in PIK3C3 (C2604T) has the potential to be a genetic marker for production traits in Duroc pigs.