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					 Physiology & ecology of longan (Dimocarpus
     longana Lour.) under acid rain stress


            Prof.Dr.Qiu Dongliang

             College of Horticulture,
Fujian Agriculture and Forestry University (FAFU)
                   P.R.China
Location of Fujian Province
 Location of Fujian Province
 Location and Climate
     FAFU is located in the western suburb of Fuzhou,the
  capital of Fujian Province and a city with a long history
  (more than 2000 years).FAFU covers an area of more
  than190 hectares.
     Fujian, a south eastern coastal province in China, with a
  populaton of 35.32 million,is affected by ocean currents
  and monsoons with a subtropical climate. The average
  annual temperature ranges between 15℃ and 22℃ with a
  rainfall between 1500~2000mm.The percentage of
  forest coverage in this province is about 63% of the
  total land mass, the highest in the nation.
One part in FAFU
          Introduction of FAFU
 The university maintains
  three postdoctoral research centers,
  one nationally assigned key discipline,
  one national center for scientific research and
  personal training,;
  6 national, top-rated doctorate programs,
  45 doctorate programs
         FAFU includes 21 colleges
   Crop Science
   Plant Protection
   Horticulture
   Forestry Science
   Life Science
   Food Science
   Economics and Business Administration
   Humanities Social Science
   Material Engineering
   Computer and information Technology
   Mechanical and Electrical Engineering
                    FAFU includes
   Resource and Environment
   Animal Science
   Bee Science
   Transportation
   Art and Landscape
   Overseas Education
   Adult Education
   Senior Professional Technique
   Jinshan
   Oriental
        Introduction of FAFU
 The 33,000 student enrollment includes
  2,500 graduate students and 23,000
  undergraduates and students seeking
  professional training courses.
Introduction of College of Horticulture,FAFU
   College of Horticulture now has a total enrollment of
1,206 full-time students, including 1,061 undergraduates
and students seeking professional training courses, 225
master students, and 25 doctoral students. The College
undertakes many projects granted by the National Natural
Sciences Foundation, the “948 Project”, the National Key
Technology R&D Program and the National Standard Plan
with funds of 20 million yuan. She is in leading position
of China in the field of sub-tropical and tropic fruit
physiology and ecology.
 Education on doctorate
  degree includes programs:
  Pomology
  Olericulture,
  Tea Science
  Ornamental Horticulture
3rd International Symposium on Longan,
    Lychee and Other Fruit Trees in
          Sapindaceae Family
    August 25~29,2008, Fuzhou,China
    (http://www.longan2008.com)
Physiology and ecology of longan (Dimocarpus
 longan Lour.) under acid rain stress includes:


  Injury of acid rain to longan and calcium regulation
   (my previous work)
  mRNA differential expression (Zheng Shan, phD
   candidate,2008)
  Special protein expression (Pan Teng-fei, phD
   candidate, 2008 )
          Previous main work

 Injury of acid rain to longan
Table 1 Injury symptom of longan under acid rain stress (Temperature(20~34℃)
───────────────────────────────────
                     pH value of simulated acid rain
 Days for stress ──────────────────────── ──
                    5.6     4.5     3.5     3.0     2.5        1.5
──────────────────────────────────
  1                 N       N       N       N        N      Brown speckles
  2                 N       N       N       N       De-greening
  3                 N       N       N       N       Brown speckles
  4                 N       N       N      De-greening
  5                 N       N       N
  6                 N       N       N
  7                 N       N       N
  8                 N       N      De-greening
───────────────────────────────────
N:No injury symptom
Table 2 Net photosynthetic rate(Pn) of longan leaves under acid rain stress
(CO2µmol/m2.s, PAR=1000µmol/m2.s)

───────────────────────

Treating pH value      Pno          Pn             Pn/Pno
───────────────────────
  5.6           7.032±1.293  7.791±1.783        1.067 a
  4.5           6.011±1.344 6.207±1.566        1.033 a
  3.5           7.307±1.976  6.637±1.363        0.908 a
  3.0           7.879±2.500  6.328±1.868        0.803 b
  2.5          6.441±0.919  4.075±0.771        0.633 c
─────────────────────--
Table 3 Effect of acid rain stress on pollen germination of longan
                   (Dimocarpus longan Lour.)


     Treatment        Pollen       Germination    Germination
                    Number(N)      Number(N)         rate

     CK1(H2O)          673             469           69.69aA
    CK2(pH 5.6)        839             590          70.32aA

       pH 4.5          827             504          60.94abA
       pH 3.5          792             365          46.09bB
       pH 2.5          815             200          24.54cC
       pH 1.5          813             78            9.59dD
Table 4 Effects of cacium spraying on Pn of longan under acid rain stress (μmolCO2.m-
2.s-1,), PAR=1000µmol/m2.s)

─────────────────────────────────
Treatment        Pno         Pn1        Pn2    Pn2/Pno Pn2/Pn1
─────────────────────────────────
 pH5.6        7.032       7.482       7.791    1.108a 1.042a
 pH3.0        7.879                   6.328    0.803c
 pH3.0+Ca      7.665      7.921       7.303    0.953b 0.922a
 pH2.5         6.441                  4.075 0.633e
 pH2.5+Ca      7.010      7.418       5.235    0.747d 0.706b
─────────────────────────────────
Ca: Concentration of Ca(NO3)2 is 15mmol·L-1
Table 5 Effect of calcium on pollen germination of longan under acid rain stress

  ───────────────────────────────
  Treatment (pH+Ca(NO3)2/mmol.L-1 ) pollen number Germination number      r/%
  ───────────────────────────────────
  4.5 + 0                            751            370                 49.27b
  4.5 + 0.02                         766           397                  51.83b
  4.5 + 0.04                         719           504                  70.10a
  4.5 + 0.06                         701           382                  54.49ab
  4.5 + 0.08                         710           360                  50.70b
  4.5 + 0.10                        732            338                 46.17b
  3.5 + 0                           714            208                 29.13b
  3.5 + 0.02                        768            272                 35.42ab
  3.5 + 0.04                        732            299                 40.85a
  3.5 + 0.06                        785            299                 38.09a
  3.5 + 0.08                        708            241                 34.04ab
  3.5 + 0.10                        688            179                 26.08b
  ─────────────────────────────────────
mRNA differential expression of
 longan under acid rain stress

 By Zheng Shan, phD candidate
 The Injury symptom of longan leaves under the
  stress of pH 2.5 of simulated acid rain.
    Partial results of DDRT-PCR primer             Reamplified cDNA from
    combination (5’T12GA 3’; 5’GATCATAGCG 3’)       differentially displayed bands
1   GATCATAGCG GCAACGGAAG CAACAACGGC TCCTTGGAGA
CAGGCAAAGC CGACAGATCA
61 GTTTGGTTGA TGAAGTGCCC GGTGGTGGTG GCTAAGTCGT
GGAAGAGTCA CGCTACTTCC
121 TCAGATCCTC AACCAGTAGC CAAGGTCGTT CTCTCTCTCG
ACCCTCTCCA ATCCGACAAT
181 ATCCAGGTAC TTCAATTTCA TTCCAGTTAT TTTAAGTTTT TTAATCTACT
CTAGTTATTG
241 TTTAAAGTAT TAGTTTTATC GTTATTCAAT TTTTGGAAAA CAAAATTCTG
TTTACGTCTC
301 CGTCGTTAAC TAGTTAGGGA TCGGTATATT TAGAGCCAGT
AGATTGATTT CTATAGCTTA
361 ATCAAGTGTT TTAGCTTTGT AATTTTAATG GCCTCTTGTT .
         Nucleotide sequence of the differential fragment T 12 GA / GATCATAGCG
TGATGAATTA TACATGTGAA/ GATCATAGCG was composed of 494 base pair .
         The fragment T 12 GA
                           The sequence TTATGAACTG TTTTCGAATT
421 TATATTATAT GTCCCACAAT of primer was underlined
TGCTTGACAA GTAGATGTTT
481 AGTG CGCTAT GATC 494
   open reading frame of the differential fragment T12GA/GATCATAGCG.
                         Blast Nucleotide




   By logging on to Genbank with the account number of FC860625, the results of BLASTN nucleotide
    sequence comparison showed that the this fragment had 75% homology with the EST of polar leaves
    under drought stress.
Special protein expression of longan
        under acid rain stress

   By Pan Teng-fei,phD candidate
SDS-PAGE in different days under stress




                A                   B
1-D electrophoresis gels analyzed by soft Quantity
                       one

                                  There was a protein band whose
                                  molecular weight is about 23KD
                                  specifically expressed under
                                  stressed by pH2.5 acid rain 15
                                  days later, and the 17KD protein
                                  specifically expressed after 5
                                  days later. Compared with pH2.5,
                                  the pH3.5 acid rain made an less
                                  influence to protein expression,
                                  but the expression differentia
                                  was raised along with the
                                  enhancement of acid rain stress.
2-dimension electrophoresis(DE)
    analyzed by PDQuest



           4-3.5       4-2.5




        4-Ca+3.5    4-Ca+2.5
                Table 6 Analysis of 2-DE(
Treatment   Up-expression   Down expression        Special expression
  CK             -                -                       -
   Ca          B1、C1              -                       C4
  2-3.5     B1、C1、E、F             A                       C4
  2-2.5     B1、C1、C3、F            A                     C4 、G
2-Ca+3.5       B1、C1              C2                      C4
2-Ca+2.5         B1               -                     C4 、G
  3-3.5        B1 、F            A、C2                    C4 、G
  3-2.5      B1 、E 、C1          A、C2          B3 、 B4 、 C4 、 C5 、 C6 、 G
3-Ca+3.5       B1 、F              A                     C4 、G
3-Ca+2.5    B1、C1、E、F             A           B3 、 B4 、 C4 、 C5 、 C6 、 G
  4-3.5      B1、E、F           A、C1 、 C2                    G
  4-2.5     B1、C1、E、F             A             B3 、 B4 、 C5 、 C6 、 G
4-Ca+3.5     B1、E、F               A                  B4 、 C4 、 G
4-Ca+2.5        E、F             A 、 B2          B3 、 B4 、 C4 、 C5 、 G
Table 7 Charactersitics of the proteins
  Protein     Iso-electric point   Molecular Weight(M)
                   (pI)
    A                6.5                 27.0KD
    B1               6.5                 59.3KD
    B2              6.45                 61.3KD
    B3              6.35                 57.3KD
    B4               6.5                 57.9KD
    C1              6.15                 59.8KD
    C2               6.0                 57.8KD
    C3              5.85                 60.0KD
    C4               6.0                 60.2KD
    C5              5.65                 58.1KD
    C6              5.65                 60.0KD
    G               6.35                 52.7KD
    E               6.64                 61.1KD
    F               6.58                 66.2KD
                  The results
 The major component of proteins were generally
  similar at different pH value acid rain, but some
  changes also took place.
  1.The 6 new specific proteins were observed.
  2.The protein C4(20.2KD pI6.0) expressed in all pH
  values acid rain during stress period.
  3. The protein F(24KD pI6.35) expressed under pH2.5
  acid rain stress in 15 days later, and then expressed in
  all pH values after 5 days later.
                  The results
4.The expression of proteins related to the acid rain pH
value and stress intensity include B3(22.3KD pI6.35),
B4(23.5KD pI6.5), C5(21.8KD pI5.6), C6(20.2KD
pI5.6).

5.The protein B4 only expressed in groups regulated
byexogenous Ca2+.
    Thanks for your attention           谢
                                        谢
                                               !
Lab of Physiology and Ecology in Horticultural Plants,
Fujian Agriculture and Forestry University,China

				
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