BackgroundSoybean pathogens and pests reduce grain production worldwide. Biotic interaction cause extensive changes in plant gene expression profile and the data produced by functional genomics studies need validation, usually done by quantitative PCR. Nevertheless, this technique relies on accurate normalization which, in turn, depends upon the proper selection of stable reference genes for each experimental condition. To date, only a few studies were performed to validate reference genes in soybean subjected to biotic stress. Here, we report reference genes validation in soybean during root-knot nematode (Meloidogyne incognita) parasitism and velvetbean caterpillar (Anticarsia gemmatalis) attack.FindingsThe expression stability of nine classical reference genes (GmCYP2, GmELF1A, GmELF1B, GmACT11, GmTUB, GmTUA5, GmG6PD, GmUBC2 and GmUBC4) was evaluated using twenty-four experimental samples including different organs, developmental stages, roots infected with M. incognita and leaves attacked by A. gemmatalis. Two different algorithms (geNorm and NormFinder) were used to determine expression stability. GmCYP2 and GmUBC4 are the most stable in different organs. Considering the developmental stages, GmELF1A and GmELF1B genes are the most stable. For spatial and temporal gene expression studies, normalization may be performed using GmUBC4, GmUBC2, GmCYP2 and GmACT11 as reference genes. Our data indicate that both GmELF1A and GmTUA5 are the most stable reference genes for data normalization obtained from soybean roots infected with M. incognita, and GmCYP2 and GmELF1A are the most stable in soybean leaves infested with A. gemmatalis.ConclusionsFuture expression studies using nematode infection and caterpilar infestation in soybean plant may utilize the reference gene sets reported here.

Seeds of the common bean, Phaseolus vulgans, contain two inhibitors of mammalian and insect a-amylases (aAls) that show specificity towards the amylases of different insect species Expression in pea (Pisum sativum) and azuki bean (Vigna angulans) of a chimeric gene consisting of the cDNA of bean aAI-1 and a seedspecific promoter makes the seeds of these legumes resistant to three species of Old World bruchids whose amylases are inhibited by aAI-1 This was the first successful genetic engineering of insect resistance in seeds To understand the basis of the specificity between amylases and inhibitors we cloned a second bean inhibitor (aAI-2) with different specificity, and we cloned the cDNA of the New World bruchid, Zabrotes subfasciatus The amylase of this bruchid is inhibited by aAI-2, but not by aAI-1 Knowledge of the ammo acid sequences and of the three-dimensional structure of the pancreatic a-amylase-aAI-1 complex allows us to predict the peptide domains and ammo acids of the proteins that are important for protein-protein recognition and inhibition of enzyme activity

Origanum vulgare L is commonly known as a wild marjoram and winter sweet which has been used in the traditional medicine due to its therapeutic effects as stimulant, anticancer, antioxidant, antibacterial, anti-inflammatory and many other diseases. A reliable gene transfer system via Agrobacterium rhizogenes and plant regeneration via hairy roots was established in O. vulgare for the first time. The frequency of induced hairy roots was different by modification of the co-cultivation medium elements after infection by Agrobacterium rhizogenes strains K599 and ATCC15834. ). The frequency of shoot generation (85.18 %) was achieved in medium fortified with 0.25 mg/L −1 of BA. Shoots were placed on MS medium with 0.25 mg/l IBA for root induction. Roots appeared and induction rate was achieved after 15 days.