First Report of Streptomyces galilaeus Associated with Common Scab in China.

During a survey of potato scab pathogens in China from 2003 to 2012, a new pathogen was found in Shanxi and Neimenggu provinces. The incidence was approximately 20% of all recovered strains. The lesions caused by the pathogen were slightly raised and similar to those caused by Streptomyces scabies (3). Lesions were excised (approximately 10 mm3) from 40 infected tubers, surface-disinfested with 0.3% NaOCl for 30 s, rinsed in sterile water three times, cut into 5 mm3, then sliced into 1-mm pieces, and plated on water agar amended with ampicillin (50 µg/ml). Plates were incubated at 28°C in the dark for 4 days. The spores of Streptomyces sp. strains growing from the tuber pieces were collected from single bacterial colonies and cultured on oatmeal agar. To fulfill Koch's postulates, one strain, CPS-2, was grown at 28°C for 10 days and the spores were washed from the plates as inoculum. One hundred milliliters of inoculum (1 × 105 CFU/ml) was mixed with autoclaved soil and vermiculite (1:1) in each pot (15 cm in diameter). Cut tubers were planted in the pots (potato cv. Favorita, one plant per pot, five replicates) and grown under greenhouse conditions (22 ± 5°C). Typical common scab symptoms consisting of small, brown, raised lesions developed on potato tubers 12 weeks after planting. The same strain was re-isolated from the lesions of the new scabby tubers. Non-inoculated plants, treated as described above, but without strain CPS-2, remained healthy. The CPS-2 strain was identified based on morphological and physiological characterization and 16S rDNA sequence. On yeast-malt extract agar, the test strain produced grayish-white aerial hypha, reddish brown substrate mycelium and pigments, and loose spiral spore chains. Spores were smooth and were 0.8 to 0.9 × 1.1 to 1.2 µm in size (diameter and length). The ability of the strain to use single sources of carbon and nitrogen was verified according to the International Streptomyces project (4). The strain grew in media supplemented with L-arabinose, D-fructose, D-glucose, rhamnose, raffinose, meso-inositol, sucrose, and D-xylose, but not D-mannitol. It used L-hydroxyproline, L-methionine, and L-histidine, and produced melanin on tyrosine and peptone yeast extract agar. The strain did not grow at a pH less than 5.0 and was sensitive to streptomycin (20 µg/ml), phenol (0.1%), and crystal violet (0.5 µg/mL), but not to penicillin (10 IU/ml). The strain also produced hydrogen sulfide. The biological characteristics of strain CPS-2 were in accord with Streptomyces galilaeus. CPS-2 produced thaxtomin A in oatmeal liquid medium and the txt AB gene fragment was successfully amplified using specific primers (2). The 16S rDNA sequence of CPS-2 was amplified by PCR with primers 16S1-F: 5'-CATTCACGGAGAGTTTGATCC-3' and 16S1-R: 5'-AGAAAGGAGGTGATCCAGCC-3' (1) and sequenced. A BLAST search of the 16S rDNA sequence for CPS-2 was conducted using the NCBI GenBank database, resulting in 99.8% similarity to S. galilaeus (NR_040857). The 16S rDNA sequence for CPS-2 (1,388 bp) was deposited in GenBank (AY621378). To our knowledge, this is the first report of S. galilaeus causing common scab of potato in China. References: (1) R. A. Bukhalid et al. Appl. Environ. Microbiol. 68:738, 2002. (2) R. Flores-González et al. Plant Pathol. 57:162, 2008. (3) D. H. Lambert and R. Loria. Int. J. Syst. Bacteriol. 39:387, 1989. (4) E. B. Shirling and D. Gottlieb. Int. J. Syst. Bacteriol. 16:313, 1966.

Inheritance and fine mapping of a restorer-of-fertility (Rf) gene for the cytoplasmic male sterility in soybean.

The cytoplasmic male sterility (CMS) line FuCMS5A and its restorer line FuHui9 were crossed to produce a segregating F(2) population for pollen fertility assay and the genetic mapping of restorer-of-fertility (Rf) gene. Results showed that the individual F(2) plants were fertile or semi-fertile based on their pollen fertility characteristics. The average ratios of viable pollen were 96.90% and 50.00% for each class of individuals. The segregation of F(2) plants showed a good fit to a 1:1 ratio, which reflects a typical heredity pattern of gametophytic CMS with fertility restorer being controlled by a single dominant gene. Using bulk segregation analysis (BSA) and genetic mapping, the Rf gene was mapped on molecular linkage group J (chromosome 16), between the simple sequence repeat (SSR) makers BARCSOYSSR-16-1064 and BARCSOYSSR-16-1082 with the distances of 0.59 and 0.83 cM, respectively. Four SSR markers (BARCSOYSSR-16-1070, Sctt011, BARCSOYSSR-16-1076 and BARCSOYSSR-16-1077) were cosegregating with this Rf gene in the mapping population. These makers will greatly facilitate the maker assisted selection procedures in CMS breeding programs and it lays a foundation for further map-base cloning of the Rf gene.

Platelet phospholipid n-3 PUFA negatively associated with plasma homocysteine in middle-aged and geriatric hyperlipaemia patients.

Studies showed that increased dietary intake of n-3 polyunsaturated fatty acids (PUFA) has a cardiovascular beneficial effect. Increased plasma phospholipid (PL) docosahexaenoic acid (22:6n-3) is associated with decreased plasma homocysteine (Hcy). The aim of this study was to investigate the relationship between platelet PL fatty acid and plasma Hcy in middle-aged and geriatric hyperlipaemia patients (50 males, 31 females) and 65 healthy subjects (43 males, 22 females) in Hangzhou, China. Plasma Hcy demonstrated significant positive correlation with adrenic acid (22:4n-6) (r = 0.188, P = 0.018) and negative correlation with 22:6n-3 (r = -0.277, P = 0.001) and the ratio of n-3/n-6 (r = -0.231, P = 0.003) in sex-, age- and BMI-controlled partial correlation analysis. The present results suggest that increased ratio of n-3/n-6 PUFA in platelet PL is associated with decreased thrombotic risks such as plasma Hcy in middle-aged and geriatric hyperlipaemia patients in Hangzhou.

[Establishment of symbiotic system for Anoectochilus roxburghii (Wall.) Lindl. and endophytic fungi].

OBJECTIVE: Setting up a symbiotic system for Anoectochilus roxburghii and endophytic fungi, so as to study the relationship between them and try to set up new cultural methods for A. roxburghii. METHOD: A. roxburghii and its endophytic fungi were cultured together on five kinds of media in flasks. The growth of the plantlets and fungi were observed. Symbiotic conditions were selected according to symbiotic characteristics. RESULTS: A symbiotic system for A. roxburghii and endophytic fungi was set up. The optimum composition of the system induded NH4NO3 825 mg.L-1, KNO3 950 mg.L-1, MgSO4 185 mg.L-1, and inositol 100 mg.L-1, other organic components being 2/3 times those of MS medium, sugrose 15 g.L-1, and other components being the same as those of MS medium, agar 9 g.L-1, pH 5.8. The culture was effected at 24-25 degrees C under cool white fluorescent light (150 lx) for a photoperiod of 11 hours. CONCLUSION: A. roxburghii and endophytic fungi can grow well together and form endomycorrhiza in the symbiotic system. Moreover, endophytic fungi help stimulate the growth and development of A. roxburghii.

Involvement of trigeminal subnucleus caudalis (medullary dorsal horn) in craniofacial nociceptive reflex activity.

We have previously shown that an increase in electromyographic (EMG) activity of digastric (DIG) and masseter (MASS) muscles can be reflexly evoked by injection into the rat's temporomandibular joint (TMJ) region of the small-fibre excitant and inflammatory irritant mustard oil (MO). Since the trigeminal (V) subnucleus caudalis (Vc, i.e. medullary dorsal horn) has traditionally been viewed as an essential brainstem relay site of nociceptive information from craniofacial tissues, an EMG study was carried out in 45 anaesthetized rats to determine if Vc is involved in the MO-evoked increases in jaw muscle EMG activity. The effects of histologically confirmed surgical or chemical lesions of Vc on this evoked EMG activity were tested in different groups of rats. MO injection into the left TMJ region of intact rats evoked bilateral increases in EMG activity of DIG and MASS which could be significantly reduced by surgical transection of the left caudal brainstem at the obex level; MO injection into the right TMJ region in these same rats still readily evoked increases in EMG activity. A sagittal section medial to Vc or transection at the level of the second cervical spinal segment did not produce any significant reduction in the reflexly evoked EMG activity. Neurones in Vc, as opposed to fibres of passage, appear to be important for the MO-evoked EMG activity, since injection into Vc of the neurotoxic chemical ibotenic acid significantly reduced the mustard oil-evoked EMG activity. The Vc also appears to play a role in the activation of contralateral V motoneurons, as evidenced by the activation of the contralateral DIG and MASS muscles by the injection of MO into the left TMJ region of intact rats and by the reduction of this evoked EMG activity in the contralateral DIG and MASS of rats with a surgical transection or ibotenic acid lesion of the left Vc. These findings suggest that Vc may be a critical element in the neural pathways underlying the reflex responses evoked bilaterally in DIG and MASS muscles by noxious stimulation of the TMJ region.

Effects of inflammatory irritant application to the rat temporomandibular joint on jaw and neck muscle activity.

An electromyographic (EMG) study was carried out in 40 anaesthetized rats to determine if the activity of jaw and neck muscles could be influenced by injection of the small-fibre excitant and inflammatory irritant mustard oil into the region of the temporomandibular joint (TMJ). Injection of a vehicle (mineral oil, 20 microliters) did not produce any significant change in EMG activity. In contrast, injection of mustard oil (20 microliters, 20%) evoked increases in EMG activity in the jaw muscles but not in the neck muscles. The increased EMG activity evoked by mustard oil was reflected in 1 or 2 phases of increased activity. The early EMG increase occurred soon after the mustard oil injection (mean latency +/- SD: 3.5 +/- 2.3 sec), peaked within 1 min, and then subsided (mean duration: 7.5 +/- 5.2 min). The later EMG increase occurred at 14.6 +/- 10.0 min after the mustard oil injection and lasted 14.3 +/- 12.3 min. These excitatory effects of mustard oil on the EMG activity of jaw muscles appear to have a reflex basis since they could be abolished by pre-administration of local anaesthetic into the TMJ region. These results document that TMJ injection of mustard oil results in a sustained and reversible activation of jaw muscles that may be related to the reported clinical occurrence of increased muscle activity associated with trauma to the TMJ.

Administration of opiate antagonist naloxone induces recurrence of increased jaw muscle activities related to inflammatory irritant application to rat temporomandibular joint region.

1. Our recent studies in rats have demonstrated that the small-fiber excitant and inflammatory irritant mustard oil injected into the temporomandibular joint (TMJ) region can evoke a sustained and reversible increase of electromyographic (EMG) activity in jaw muscles and an acute inflammatory response. The aim of the present study was to test if opioid mechanisms are involved in modulating the EMG increase evoked by mustard oil. 2. Mustard oil injected into the rat TMJ region evoked significant increases of jaw muscle EMG activity; the vehicle mineral oil had no such effect. The increased EMG activity lasted up to 20 min, and by 30 min after the mustard oil injection had returned to control (preinjection) levels, at which time administration of the opiate antagonist naloxone (1.3 mg/kg i.v.) induced a significant recurrence of the increase in EMG activity. This "rekindling" of EMG activity appeared at 5 to 10 min after the naloxone administration and lasted for 10 to 20 min. In contrast, naloxone administration in the animals receiving mineral oil injection into the TMJ region did not "rekindle" the EMG activity, nor did the administration of the peripherally acting opiate antagonist methylnaloxone or the vehicle of naloxone. 3. These findings reveal that the application of the opiate antagonist naloxone produces a recurrence of increased jaw muscle activity reflexively evoked by mustard oil injection into the rat TMJ region. They suggest that central opioid depressive mechanisms activated by the mustard oil-induced afferent barrage limit the duration of the evoked EMG changes.

Excitatory effects on neck and jaw muscle activity of inflammatory irritant applied to cervical paraspinal tissues.

A study was carried out in 19 anaesthetized rats to determine if the electromyographic (EMG) activity of jaw and neck muscles could be influenced by injection of the inflammatory irritant mustard oil into deep paraspinal tissues surrounding the C1-3 vertebrae. The EMG activity was recorded ipsilaterally in the digastric, masseter and trapezius muscles and bilaterally in deep neck muscles (rectus capitis posterior). In comparison with control (vehicle) injections, mustard oil (20 microliters, 20%) injected into the deep paraspinal tissues induced significant increases in EMG activity in the neck muscles in all the animals and in the jaw muscles in the majority of the animals; the effects of mustard oil were more prominent in the former. The EMG response evoked by mustard oil injection was frequently reflected in two phases of enhanced activity. The early phase of the increase in EMG activity was usually initiated immediately following mustard oil injection (mean latency: 20.4 +/- 17.7 sec) and lasted 1.6 +/- 1.1 min. The second phase occurred 11.3 +/- 7.6 min later and lasted 11.0 +/- 8.1 min. Evans Blue extravasation was apparent in the deep paraspinal tissues surrounding the C1-3 vertebrae after mustard oil injection, and histological examination showed that mustard oil injection induced an inflammatory reaction in the rectus capitis posterior muscle. These results document that injection of the inflammatory irritant mustard oil into deep paraspinal tissues results in a sustained and reversible activation of both jaw and neck muscles. Such effects may be related to the reported clinical occurrence of increased muscle activity associated with trauma to deep tissues.

Differential effects of cutaneous and deep application of inflammatory irritant on mechanoreceptive field properties of trigeminal brain stem nociceptive neurons.

1. The aim of this study was to determine whether there are differential neuroplastic effects of cutaneous and deep stimuli on the mechanoreceptive field (RF) properties of nociceptive brain stem neurons recorded in trigeminal (V) subnucleus caudalis. 2. A total of 31 caudalis neurons were functionally identified in anesthetized rats as nociceptive neurons receiving convergent cutaneous and deep afferent inputs, and the effects of the small-fiber excitant and inflammatory irritant mustard oil applied to either tongue muscle or facial skin were tested on their cutaneous and deep RFs. 3. The expression of neuroplasticity on the cutaneous and deep RFs of the neurons was found to depend on the site of application of this irritant. Mustard oil injected into the tongue muscle produced a significant and reversible expansion of both the cutaneous and deep RFs. In contrast, application of mustard oil to facial skin resulted in a significant and reversible expansion of the cutaneous RF, but no deep RF expansion occurred. 4. These findings suggest that considerable functional neuroplasticity of the cutaneous and deep RF properties of V nociceptive neurons can be induced by a peripheral inflammatory irritant and that deep inputs are especially effective in evoking these neuroplastic changes.

Biosynthesis of vasopressin by gastrointestinal cells of Brattleboro and Long-Evans rats.

The distribution of vasopressin, provasopressin, vasopressin-associated neurophysin, and vasopressin-associated glycopeptide was determined immunohistochemically in the gastrointestinal system of Brattleboro and Long-Evans rats. Cells containing immunoreactivity for vasopressin, provasopressin, neurophysin, and glycopeptide were detected in the same cell types of the stomach and duodenum, while selected cells in the duodenum contained only immunoreactive glycopeptide. Unlike that in the hypothalamus, staining for neurophysin in the gastrointestinal tract was sensitive to fixation. These findings indicate that vasopressin is produced by cells in the rat gastrointestinal system and suggest the existence of synthetic pathways different from those found in hypothalamic neurons.

[The oculocardiac reflex during ocular operation under various anesthesia].

100 patients underwent extraocular muscle tugging of 154 extraocular muscles during ocular operations to study the oculocardiac reflex. The results showed that retrobulbar block was effective in minimizing occurrence of the reflex whether under general or local anesthesia.