D E C E M B E R 2021

SUN MON TUE WED THU FRI SAT

01 02 03 04

05 06 07 08 09 10 11

12 13 14 15 16 17 18
: Date I created this : Prepare soil mix (7,000 : Prepare soil mix with
calendar lol kg LS + 3,500 kg SAND) Copper Sulfate at 10AM
: Check soil conditions

19
: Extract seeds from seed 20 21 22 23 24 25
coat (1st) at 7 AM
: Pretreatment (2nd) and : Check and record : Check and record
sowing at 10 AM humidity & temp humidity & temp
: Check humid & temp. & : Sown 2 seeds with : Sown ALL SEEDS;
soil control treatment experiment proper

26 27 28 germinated 29 30 31
Failed to
germinate 2 1st 2nd
seeds
 J A N U A R Y 2022
SUN MON TUE WED THU FRI SAT
01
3rd

02 03 04 05 06 8th (First count) 07 08

4th 5th 6th 7th : Check and record
humidity & temp
: Sown seeds FIRST
COUNT

09 10 11 12 13 14 15

17 18 19 20 21 22 23

24 25 26 27 28 29 30

31

F E B R U A R Y 2022
 SUN MON TUE WED THU FRI SAT
01 02 03 04 05

06 07 08 09 10 11 12

13 45TH DAY 14 15 16 17 18 19
Transplant seedlings Conduction 2.0
– measure shoot -Cracking and
And root length sowing seed

20 21 22 23 24 25 26

27 28
 M A R C H 2022
SUN MON TUE WED THU FRI SAT
01 02 03 04 05

06 07 08 09 10 11 12
Start of 2nd sem, 1st
cycle

13 14 15 16 17 18 19

20 21 22 23 24 25 26

27 28 29 30 31

I. Collection of Seeds
 The Z. talanai seeds were obtained from Ecosystem Research
and Development Bureau (ERDB) at Los Baños, Laguna. Taken into
consideration the ecological status and availability, a total
of 120 seeds were used, as modified from study of Chen et al.
(2012). To maintain good condition for the seeds, ERDB
instructed wrapping them with cloth and refrigerate until
sowing day.
II. Preparation of Experimental Treatments
The optimal sowing medium for Ziziphus is sandy-loam soil
(Hussein et al., 1996; Danin, 2013).

Here, sandy-loam soil samples were sterilized using oven for

60 minutes at a temperature of 100 oC (Raducanu et al., 2016).

The sterilized soil was artificially contaminated with

copper in a technical grade copper sulphate pentahydrate
(CuSO4.5H2O) form.

Different concentrations of CuSO4.5H2O were based on the

reference set by Environmental Company of the State of São
Paulo (2016): Cu contamination in soil starts with Prevention
value of 60 mg kg-1; Agricultural Intervention of 760 mg kg-1;
Residential Value of 2,100 mg kg-1; and Industrial
Intervention Value of 10,000 mg kg-1. After thorough mixing,
the soil samples were acclimatized for 24 hours with the
CuSO4.5H2O (Wyszkowska et al., 2017).

III. Extraction and Pre-treatment of Seeds

Prior to sowing, Ziziphus seeds undergone pre-treatment to
overcome seed mechanical dormancy (Schmidt, 2000). The
cracking of the hard endocarp of the seeds had been utilized
and recorded for efficiency in breaking the dormancy of other
Ziziphus species’ seeds (Dorji, et al., 2016; Saied et al.,
2008). After three (3) weeks of storage, the Z. talanai seeds
were cracked open from endocarp, extracting two seeds. The
extracted seeds with healthy, brown seed coat with no
observable damaged after cracking were used for the
experiment.
IV. Germination Experiment
The pre-treated Z. talanai seeds were sown in inverted seed
cells of plastic trays (Nejat and Sadeghi, 2016), containing
the soil treatments in 1.5 cm burial depth (Maraghni et al.,
2010).
The treatment groups: Sandy loam soil + 0 Cu (CuSO4.5H2O)
ppm, Sandy loam soil + 60 Cu (CuSO4.5H2O) ppm, Sandy loam soil
+ 760 Cu (CuSO4.5H2O) ppm, Sandy loam soil + 2,100 Cu
(CuSO4.5H2O) ppm, and Sandy loam soil + 10,000 Cu (CuSO4.5H2O)
ppm were given eight (8) seeds for each replication.
The seed were placed in a greenhouse tent, with monitored
60% (+10%) relative humidity (Takhti and Shekafandeh, 2012),
and temperature range of 25 – 40 oC, as from the collective
information on other Ziziphus species reports of ideal
temperature for germination (Maraghni, et al., 2010; Aráoz et
al., 2004; Laamouri and Zine El Abidine 2000; Danthu et al.,
1993; Speer and Wright 1981).
The seeds were exposed to 12 hours light and 12 hours dark
(Dorji et al., 2015) and received moisture with distilled
water periodically (Saqueira et al., 2020).
The seeds were reported to germinate once the radicle
exceeds 2 mm in length (Saqueira et al., 2020). Here, eight
(8) days after sowing was considered the start of the first
count until 15th day as the final count of germinated seeds
(Del Longo and Aráoz, 2009).
The experiment continued until the 45th day (Takhti and
Shekafandeh, 2012; Saqueira et al., 2020) to keep the
observation of establishment status. Data and measurements
were recorded daily.

Reference:
Raducanu, Dumitra & Stefanescu, Ioana & Ifrim, Irina &
PATRICIU, Oana-Irina & Rati, Ioan & Prisecaru, Maria &
Stamate, Marius & Iuliana, Lazar. (2016). Effects of different
sanitization treatments on soil characteristics. Environmental
Engineering and Management Journal. 15. 624-634.
10.30638/eemj.2016.068.