|
| Status |
Public on Feb 01, 2007 |
| Title |
Dodder_Pumpkin_Rep2 |
| Sample type |
RNA |
| |
|
| Source name |
Dodder grown on pumpkin replicate 2
|
| Organism |
Cuscuta pentagona |
| Characteristics |
Cuscuta pentagona
|
| Biomaterial provider |
Dr. Tom Lanini, University of California, Davis
|
| Treatment protocol |
Lespedeza dodder (Cuscuta pentagona) seeds were scarified by soaking in concentrated sulfuric acid for one hour, rinsed thoroughly in water, and planted around the base of two-week old tomato plants. Dodder stems were trained from tomato to four-week old pumpkin plants and dodder connections between tomato and pumpkin were severed two weeks after the formation of new haustorial connections with pumpkin. Dodder stems at least 4 cm from the host plant were harvested after 9-12 weeks of growth. To obtain sufficient quantity of dodder tissue, many stems were harvested, representing multiple individuals for each harvest time. All tissues were immediately frozen in liquid nitrogen and stored at -80ºC.
|
| Growth protocol |
Pumpkin (Curcurbita maxima, Blue Hubbard Winter Squash, Heirloom Seeds, West Elizabeth, PA) plants were grown in Metro Mix 360 potting medium (Griffin Greenhouse and Nursery Supplies, Richmond, VA) under greenhouse conditions.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA (1-1.5 g) was extracted from dodder by grinding in liquid nitrogen, and extracted using TRIzol reagent (Invitrogen, Carlsbad, CA). The RNA was precipitated with 100% isopropanol followed by a second precipitation with 100% ethanol and 3M sodium acetate (pH 5.2). The RNA pellet was dissolved in DEPC-treated water and treated with DNase using a DNA-free Kit (Ambion, Austin, TX).
|
| Label |
Biotin
|
| Label protocol |
A linear amplification of total RNA was performed using Ovation Biotin RNA Amplification and Labeling System (NuGEN, San Carlos, CA).
|
| |
|
| Hybridization protocol |
Hybridization was conducted according to Affymetrix standard procedure and performed by the Virginia Bioinformatics Institute Core Laboratory.
|
| Scan protocol |
Scanning was conducted according to Affymetrix standard procedure and performed by the Virginia Bioinformatics Institute Core Laboratory.
|
| Description |
The objective of this experiment was to assay for movement of mRNA from tomato (host) into dodder (parasitic plant). Dodder was grown on pumpkin as one of several control hosts to facilitate identification of endogenous dodder mRNA from those potentially translocated from the .
|
| Data processing |
Data was not processed beyond the values provided by the Affymetrix intensity values.
|
| |
|
| Submission date |
Jan 12, 2007 |
| Last update date |
Jan 17, 2007 |
| Contact name |
James H Westwood |
| E-mail(s) |
westwood@vt.edu
|
| Phone |
540-231-7519
|
| Fax |
540-231-3347
|
| Organization name |
Virginia Tech
|
| Department |
Plant Pathol. Physiol. Weed Sci.
|
| Street address |
401 Latham Hall
|
| City |
Blacksburg |
| State/province |
VA |
| ZIP/Postal code |
24061-0390 |
| Country |
USA |
| |
|
| Platform ID |
GPL4741 |
| Series (1) |
| GSE6736 |
Trafficking of mRNA from tomato to dodder |
|
