WORCESTER POLYTECHNIC INSTITUTE
The long-term goal of the current project (2 R15 GM069562-02) is to develop a fundamental understanding of the regulatory control of terpene production in Artemisia annua so that in planta metabolic manipulation of terpenoid biosynthesis can eventually be developed not only for artemisinin (AN), but also other therapeutically important plant produced terpenoid drugs. The current work has 3 objectives: 1) To compare the relative expression of the early (DXS, DXR, HMGR, and FPS) and late (ADS and CYP71AV1) genes in terpenoid biosynthesis with levels of both AA and AN production in A. annua seedlings as they are shifted from vegetative growth to conditions that induce flowering.
With supplemental funds we proposed doing the following that are encompassed within the original IRG-approved scope as part of our original Objective #1 (see above):
1. Use LC/MS to compare AN and AN metabolites (artemisinic acid, dihydroartemisinic acid and AN) production in shoots that begin to develop roots in vegetative plants. This is directly linked to Objective # 1 because it enables us to compare unrooted shoots to rooted shoots during the vegetative stage of growth. This was not possible until little over a year ago when we obtained a reliable, high AN producing clone of A. annua that can be cultured in vitro as either shoots or as rooted shoots. We have been culturing this clone for over a year and using it in other experiments aimed at overproduction of AN, but not related to a fundamental study of developmental regulation.
2. We will further analyze the transcript levels of the genes in the AN biosynthetic pathway (identified in the original IRG-approved scope) and establish correlations between those genes that may get turned on or off at the level of transcription as the roots develop during the vegetative growth stage and especially while the roots are being initiated.
Together these additional data will enable us to better correlate artemisinin biosynthesis with the development of roots during the vegetative developmental phase of A. annua plant growth thereby better enabling completion of our original Aims.
| AWARD OVERVIEW |
| Award Number |
3R15GM069562-02S1 |
Funding Agency |
Department of Health and Human Services |
| Total Award Amount |
$69,362 |
Project Location - City |
Worcester |
| Award Date |
09/25/2009 |
Project Location - State |
MA |
| Project Status |
Completed |
Project Location - Zip |
01609-2280
|
| Jobs Reported |
0.08 |
Congressional District |
03 |
| Project Location - Country |
US |
|
|
Recipient Information
(Grants)
| Recipient Information (Grants) |
|
Recipient Name
|
WORCESTER POLYTECHNIC INSTITUTE |
| Recipient DUNS Number |
041508581
|
| Recipient Address |
100 INSTITUTE RD |
| Recipient City |
WORCESTER |
| Recipient State |
Massachusetts |
| Recipient Zip |
01609-2280 |
| Recipient Congressional District |
03 |
| Recipient Country |
USA |
Required to Report Top 5
Highly Compensated Officials |
No |
Projects and Jobs Information
| Projects and Jobs Information |
| Project Title |
Artemisinin Biosynthesis: Control of Transcription |
| Project Status |
Completed |
| Final Project Report Submitted |
Yes |
| Project Activities Description |
Science & Technology, General/Other |
| Quarterly Activities/Project Description |
We slightly altered our experimental approach to better address the relationship between rooting and AN production in A. annua. Ms. Nguyen completed experiments to determine if the medium used to initiate shoots will halt AN formation in rooted shoots. After 12 days incubation in shooting or in rooting medium, rooted shoots did not show any significant difference in AN levels. This result suggested that once roots are present, then the signal to make AN is ensconced and irreversible. When analyzing the metabolites for this project, we found it was more important to measure AN, arteannuin B (AB), and deoxyAN instead of artemisinic acid (AA) and dihydroartemisinic acid (DHAA) because those 3 are the end products of what we have now recently identified as 3 branches of the pathway (unpublished). Only AN was detectable in the leaves and, as previously reported (Mannan et al., 2010), increased significantly by > 10 fold over that of unrooted shoots. Interestingly, no AB or deoxyAN was detected suggesting that at least under in vitro conditions, and during rooting, these pathways may be less active. Compared to unrooted shoots, transcripts of ADS and CYP, increased slightly, but only after roots formed after day 8 (Fig. 1). Although not part of our Aims, we also included trichome populations in our measurements. As the roots formed on cultured shoots, the number of trichomes appeared to decrease, but their size seemed to increase; unfortunately, none of these changes was statistically significant. Together these data show there is no measureable trichome change with rooting, but there is some response in ADS and CYP transcripts, which correlates well with the observed increase in AN. This student will continue this work to determine if by increasing the number of replicates, a statistically significant set of data can be obtained. |
| Jobs Created |
0.08 |
| Description of Jobs Created |
Two undergraduates, Andrew Keyser and Brant Carter, were hired part time to help establish clonal lines of the PEG01 Chinese version of Artemsia annua. Six lines are now successfully established, tested, and their use in experiments has already proved successful; they greatly reduced population variability of our experiments, so fewer replicates are required.
|
Purchaser Information
(Grants)
| Purchaser Information |
| Contracting Office ID |
Not Reported |
| Contracting Office Name |
Not Available |
| Contracting Office Region |
Not Available |
| TAS Major Program |
75-0852 |
| Award Information |
| Award Date |
09/25/2009 |
| Award Number |
3R15GM069562-02S1 |
| Order Number |
|
| Award Type |
Grants |
| Funding Agency ID |
75 |
| Funding Agency Name |
Department of Health and Human Services |
| Funding Office Name |
Not Available |
| Awarding Agency ID |
75 |
| Awarding Agency Name |
Department of Health and Human Services |
| Amount of Award |
$69,362 |
| Funds Invoiced/Received |
$69,362 |
| Expenditure Amount |
$69,362 |
| Infrastructure Expenditure Amount |
$0 |
| Infrastructure Purpose and Rationale |
Not Reported |
| Infrastructure Point of Contact Name |
Not Reported |
| Infrastructure Point of Contact Email |
Not Reported |
| Infrastructure Point of Contact Phone |
Not Reported |
| Infrastructure Point of Contact Address |
Not Reported |
| Infrastructure Point of Contact City |
Not Reported |
| Infrastructure Point of Contact State |
Not Reported |
| Infrastructure Point of Contact Zip |
Not Reported |
Product or Service Information
(Grants)
| Product or Service Information |
| Primary Activity Code |
U01 |
| Activity Description |
Science & Technology, General/Other |
| Sub-Awards Information |
| Sub-awards to Organizations |
0 |
| Sub-award Amounts to Organizations |
$0 |
| Sub-Awards to Individuals |
0 |
| Sub-Award Amounts to Individuals |
$0 |
| Number of Sub-awards less than $25,000/award |
0 |
| Amount of Sub-awards less than $25,000/award |
$0 |
| Number of payments to vendors greater than $25,000 |
0 |
| Total Amount of payments to vendors greater than $25,000/award |
$0 |
| Number of payments to vendors less than $25,000/award |
0 |
| Total Amount of payments to vendors less than $25,000/award |
$0 |
| Location Information |
| Latitude, Longitude |
42º 16' 21",
-71º 48' 27" |
| Congressional District |
03 |
| Address 1 |
100 Institute Rd |
| Address 2 |
|
| City |
Worcester |
| County |
Worcester |
| State |
MA |
| Zip |
01609-2280 |
|
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