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Jose Judy Stervil (1), Barbara Hyacinthe (1), Dora Pilar Maul (1), Barbara Freeman (2) David Kuhn(2).
(1) School of Science, St. Thomas University, 16401 NW 37th Avenue, Miami Gardens, FL 33054, (2) USDA-ARS-SHRS, 13601 Old Cutler Road, Miami, FL 33158 .
I.Introduction
III. Results
Mango (Mangifera indica) is a highly consumed tropical crop because of its unique
1. RNA extraction and quantification of Mango
flavor, color, and shape. It belongs to the Anacardiaceae family of the flowering plants
and is native to South and Southeast Asia. There are four genetically diverse groups of
mango: Indian, Floridian, Southeast Asian, and West Indies (Schnell et al., 2006)
Due to the great demands of commercially favored mangoes, the United States
Department of Agriculture-Agricultural Research Service - Subtropical Horticulture
Research Laboratory (USDA-ARS-SHRL) in Miami, FL has established several
research projects among which are Mango SNP discovery and gene expression in
developing mango fruit. Both projects involve RNA extraction steps. After RNA
Sequencing, sequences can be used to detect Single-Nucleotide Polymorphism (SNPs),
which can be used as markers to create a genetic map in order to determine the location
of the mutations in the chromosomes. Whether a gene is expressed can also be
identified using the extracted RNA combined with other techniques. Mango phenotypic
data can be compared to the genetic map in order to link phenotypic traits to genetic
Figure 1. The samples were
Figure 2. Chloroform was used to
markers.
pulverized using liquid nitrogen
separate the RNA from the rest of
In this experiment, RNA was isolated from four stages of developing fruit, which
the organic compounds. The RNA
during the extraction of RNA.
remained in the top layer.
includes exocarp, mesocarp, seed coat and seed. In addition, phenotypic data of various
mango cultivars harvested from the station were collected for a period of 4 weeks.
Objectives of the present experiment:
Extract RNA to be used for SNP discovery and gene expression
2. Collection of Mango Phenotypic Data
Collect the phenotypic data for several cultivars of mangoes from the USDA-ARS
station
V. Future Steps
Location
RINe
E1
4.3
28S/18S
Conc. [ng/l] Sample Description
(height)
0.7
34.0
29_1-2seedMauri
A1
4.1
0.4
2.89
TA47_4-3scoatTAtkins
B1
4.0
0.3
155
TA48_4-3seedTAtkins
C1
6.4
1.9
1670
27_1-2exocarpMauri
D1
7.2
1.5
650
28_1-2mesocarpMauri
F1
7.4
0.8
112
30_1-3exocarpMauri
G1
6.3
1.0
142
31_1-3mesocarpMauri
H1
A2
5.3
5.0
0.7
1.8
6.85
315
32_1-3seedMauri
33_2-2exocarpMauri
B2
8.3
1.3
174
34_2-2mesocarpMauri
C2
D2
A0
4.4
3.0
1.3
0.9
122
30.9
33.3
Pot1_PI545851
Pot2_PI545851
ladder
VI. Acknowledgements
The authors want to acknowledge
Ashley Johnson, Leah Schwartz,
Herma Pierre, Tomas AyalaSilva. A special thanks to Carlos
Vazquez for his support. Partial
funding for this project came the
USDA-HIS-funded FCCAgE
Agricultural Education grant and
from the U.S. Department of
Education - STEM TRAC
grant(P03C110190), MDC
School of Science-STU School
of Science, Technology and
Engineering management.
VII. References
Bailey et al., 2005. Gene
expression in leaves of Theobroma
cacao in response to mechanical
wounding, ethylene, and/or methyl
jasmonate. Plant Science 168,
1247-1258.
Kuhn et al., 2012. Identification
and mapping of conserved ortholog
set (COS) II sequences of cacao
and their conversion to SNP
markers for marker-assisted
selection in Theobroma cacao and
comparative genomics studies.
Tree Genetics & Genomes 8, 97
111.
Schnell et al., 2006. Mango
Genetic Diversity Analysis and
Pedigree Interferences for Florida
Cultivars Using Microsatellite
Markers. J. Amer. Soc. Hort. Sci.
131(2), 214-224.