Detection of GLV, OYDV and LYSV in potato onion (Allium cepa L., Aggregatum group) by RT-PCR

Three pairs of primers were designed and synthesized from nucleotide sequences of garlic latent virus （GLV）, onion yellow dwarf virus （OYDV）, and leek yellow stripe virus （LYSV） by using PCR primer design software. The expected fragments about 170 bp, 287 bp, and 191 bp were amplified by RT-PCR for GLV, OYDV, and LYSV, respectively in disease-infected plants of potato onion （Allium cepa L., Aggregatum group）, but such fragments were not obtained from healthy-looking plants and virus-free seedlings of shoot-tips. The amplified products ofGLV, OYDV and LYSV were cloned into pGEM-T vectors, and transformed into Escherichia coli. JM109. The recombinant plasmids were obtained and sequenced. The nucleotide sequences were compared with corresponding viral nucleotide sequences reported in GenBank by performing a NCBI BLAST. The analysis showed that their homology attained 75% to 90%,89.5% to 96.1%,and 91.6% to 96.3% in GLV, OYDV, and LYSV, respectively. The total RNA of 6.34 ug·uL^-1 from infected plants was diluted to a series of 10^-1 to 10^5 and the detection sensitivity of RT-PCR was 10^4 （about 4 ng）. Thus, a method of identification and detection by RT-PCR of GLV, OYDV, and SLYV was established.

EMS-Mediated Mutagenesis in Marigold Seeds and Its Effects on Seedling Growth and Physiology

Marigolds(Tagetes spp.)are popular horticultural plants worldwide.The current study aimed to investigate the optimal mutagenic conditions for marigold seeds using EMS(ethyl methanesulfonate)mutagenesis.Different con-centrations and treatment times of EMS were applied to investigate their effects on the marigold seed germination rate,growth traits,antioxidant enzyme activities(i.e.,SOD and POD),and malondialdehyde(MDA)contents.Results indicated that with increasing the EMS treatment duration and concentration,the seed germination rate and growth treatments were reduced,accompanied by elevated MDA content.In addition,SOD and POD activ-ities initially correlated positively with the growth tratis at the lowest concentrations and shortest durations of EMS,but such relationship diminished beyond certain thresholds.The comprehensive analysis identified the opti-mal mutagenic conditions as 1%EMS treatment for 12 h,achieving a semi-lethal dose and enhancing stress-resis-tant components in seedlings.Thesefindings are pivotal for advancing genetic enhancement and germplasm innovation in marigolds.

Seasonal occurrence of Aphis glycines and physiological responses of soybean plants to its feeding

The soybean aphid Aphis glycines Matsumura （Hemiptera： Aphididae） is an important pest of soybean in China. To monitor and manage this pest effectively it is neces-sary to understand its population dynamics and demographics, as well as the physiological responses of soybean plants to its feeding. In this study, using field surveying and suction-trap monitoring, we investigated the population dynamics of the soybean aphid in Xiuyan County, Liaoning Province in northeastern China during 2009-2012. The results indicatedthat the population dynamics of the soybean aphid followed a unimodal curve distribution, with the insect generally colonizing soybean fields from the middle of June to early Julyand the population reaching a peak between early July and early August. On the whole, soybean aphids occurred in suction-traps at least 2 weeks earlier than they were foundin field surveys. A total of 72 alates were collected by suction-trapping over the 4 years, with the earliest alate captures occurring on 28 May in 2009, 2011, 2012 and 4 June in 2010. The life table parameters clearly showed that this aphid had a short doubling time （4.73 ± 0.21 days）, and 7.36± 0.98 nymphs were produced by a soybean aphid adult during its lifetime （13.57 ± 0.30 days）. Finally, biochemical assays indicated that the amount of malondialdehyde and the activities of four defense-related enzymes in soybean leavessignificantly changed between 0 day and 7 days of aphid infestation. Polyphenol oxidase （PPO） and catalase （CAT） activities increased more dramatically after 1 day of aphid feed-ing. In addition, significantly higher levels of superoxide dismutase and CAT were found after aphid feeding for 7 days, whereas there was no significant change in the activitiesof peroxidase and PPO. Consequently, this study will be beneficial in determining the seasonal occurrence of the soybean aphid and selecting insect-resistant soybean varieties,and thus in developing a theoretical framework for appropriate management strategies.

Artificial Shiro formation of Tricholoma matsutake

One type of soil collected from Maoer-shan in Heilongjiang Province,China was selected to induce hyphal growth of Tricholoma matsutake by a soil screening ex-periment.It was confirmed that hyphal growth of all the tested T.matsutake isolates was significantly stimulated in soil by supplemented with 0.5%~2.0%olive oil.The ag-gregation of hyphae and soil resembled natural Shiro.The biomass of hyphae in the soil increases with increasing olive oil concentrations.Moreover,seedlings of Pinus den-siflora grew well in the soil containing 0.5%~1.0%olive oil and were also successfully infected by T.matsutake isolate A in the soil containing 1.0%olive oil.This study estab-lished a culture system of artificial Shiro formation and also provided a premise for formulation of culture substratum for fruit body formation of T.matsutake.